Through a range of methods we have thus contributed an empirically grounded and theoretically

informed understanding of climate vulnerability. With our seasonal calendars, explicitly building on our field data and design, we are able to study the temporal interactions between nature and society, thereby considering climatic, agronomic and disease dynamics in a place-based setting, as suggested by Thompson (2009). From this we show that time and timing are significant for understanding exposure, sensitivity and adaptive capacities in any attempt to contextualize climate vulnerability. Not only does this exercise generate insights into how these stressors are interrelated, i.e., how they feed into and off each other by contributing to different sensitivities at different times of the year, depending on the type of exposure, it also illustrates that when exposure, sensitivity and limited adaptive capacity converge BIX 1294 in vivo in time, climate vulnerabilities are greater because of destructive reinforcing feedbacks check details on the human-environment system. In addition, we show that farmers engage in continuous, yet reactive and autonomous adaptation to climate vulnerability by relying on past experiences of dealing with climate extremes, despite their waning viability in times of increasing climate uncertainty. Current differential adaptive capacities between households and communities

indicate Bay 11-7085 a deficit in adaptation potential among smallholder farmers in the LVB, which makes life especially troublesome and the future highly uncertain. In all this, age and gender are pronounced aspects of the capacity of a person, a household or a community to

cope with climate-induced impacts, not to mention increasing the adaptive capacities to reduce climate vulnerability. The wheel of hardship underscores how households rely on a selleck inhibitor steady flow of cash, food and (healthy) labor power to manage converging aspects of exposure and sensitivities. Historically, farmers have often managed this through increased diversification, which is also seen as a strategy emphasized and promoted by the World Bank (2008). However, our study illustrates that livelihood diversification at household levels is becoming increasingly undermined as a livelihood strategy and that the alternatives, in terms of migration and extension of agriculture, now offer only limited opportunities. The only other feasible adaptation strategy for the LVB is therefore to intensify agricultural production. But, as previously mentioned, this hinges not only on peoples’ ability to pool labor but also on increased knowledge about how to farm more sustainably in times of global environmental change (Pretty et al. 2011). To enable farmers to do this clearly requires governmental action and financial investment.

Degradation of trehalose-6-phosphate can be mediated by a Pritelivir clinical trial trehalose 6-phosphate hydrolase (TreC), belonging to family 13 of glycoside hydrolases [16], or a trehalose-6-phosphate phosphorylase (TrePP) [19].Trehalase, trehalose phosphorylase, and trehalose-6-phosphate Doramapimod datasheet hydrolase were detected in soybean nodules formed by B. japonicum[20], but orthologous genes for these enzymes were not found in the genome of S. meliloti[21]. In the former species, two ABC transport systems (ThuEFGK and AglEFGAK), and one major catabolic pathway (ThuAB) have been reported for trehalose [22, 23]. In rhizobia, the effect of trehalose

accumulation on tolerance to osmotic and drought stress, as well as symbiotic performance, appears to be dependent on the particular stress, the rhizobial species, and the host genotype. Regarding osmotic stress, OtsAB seems to play a major role in trehalose accumulation under hyperosmotic conditions, click here and it is the main system involved in osmoadaptation of S. meliloti[5] and B. japonicum[2]. In addition, accumulated trehalose seems to have

a major role in protecting B. japonicum[24] and R. leguminosarum bv trifolii[7] against desiccation stress. With respect to symbiotic phenotype, in B. japonicum trehalose accumulation is involved in the development of symbiotic nitrogen-fixing root nodules on soybean plants [2]. In contrast, in other rhizobia such as R. leguminosarum bv trifolii or S. meliloti, trehalose accumulation has been proposed to be important

only for competitiveness [5, 7]. The role of trehalose as thermoprotectant has been established in yeast [25] and bacteria such as E. coli[26], Salmonella enterica serovar Typhimurium [27] or the halophilic bacterium Chromohalobacter salexigens[28]. However the role of trehalose in protection against heat stress in rhizobia has not yet been investigated. Common bean (Phaseolus vulgaris) is an important crop in the diet of people 4��8C of Latin America. In this region, it is mainly nodulated by R. etli[29]. The complete genome sequence of R. etli CFN 42 has been reported ( http://​www.​ccg.​unam.​mx/​retlidb/​) [30]. It contains more replicons (a circular chromosome and six large plasmids) than any other completely sequenced nitrogen-fixing bacterium, but several pieces of evidence suggest an exogenous origin for plasmids p42a and p42d Suarez and co-workers [10] reported an otsA mutant still capable of accumulating trehalose to a certain extent, which was nevertheless osmosensitive and displayed reduced nodulation and lower nitrogenase activity, and consequently reduced plan biomass. In contrast, an OtsA overexpressing R. etli strain showed increased trehalose content and was more tolerant to osmotic stress than the wild-type. Bean plants inoculated with the OtsA overexpressing strain showed improved nodulation and nitrogen fixation, and increased drought tolerance.

Interestingly, the cells harbouring the two AidB-YFP foci are significantly (p < 0.005) smaller

Fludarabine (1.93 μm on average) than the bacteria having a single focus of AidB-YFP at the constriction site (2.08 μm on average), suggesting that in the cell cycle, bacteria with 2 foci precede those with a single focus at the constriction site (Figure 3A). This feature of the cell cycle is depicted in the discussion. Figure 3 Size distribution of B. abortus carrying AidB-YFP, in the presence or absence of an alkylating agent (EMS). The bacterial lengths were grouped in classes of 0.25 μm, and the maximum value for each class is given on the × axis. (A) Size distribution of 276 bacteria (XDB1128 strain) with AidB-YFP either at the new pole (white), the new pole and the constriction site (dark grey), or the constriction site only (black). (B) Size distribution of B. abortus (XDB1128 strain) exposed to 0.4% of EMS for 4 h (light grey, n = 340) or the unexposed control (white, n = 218, bacteria without detectable constriction). selleck inhibitor (C) DIC and fluorescence pictures of the XDB1128 strain expressing aidB-yfp and pdhS-mCherry fusions, as described in figure 2. The bacteria in the lower panels have been exposed to 0.4% EMS for 4 h in rich (2YT) medium. On the

top panels, control bacteria were incubated for 4 h in 2YT in the absence of EMS. Constriction sites are indicated by arrowheads. Each scale bar represents 2 μm. Furthermore, the localization of AidB-YFP is still at the new pole after 4 h of exposure with 0.4% EMS (80% of the bacteria exhibited PdhS-mCherry at one pole and AidB-YFP at the opposite pole, n = 237). This selleck screening library observation indicated that AidB-YFP is not released from the new pole in the presence of an alkylating stress with EMS, further suggesting that AidB is active at the new pole, because in these conditions an aidB mutant is killed. Interestingly, bacteria exposed to EMS displayed detectable constriction at the much less frequency (2 constrictions observed among 254 bacteria) compared to the

untreated control (44 constrictions observed among 254 bacteria). Moreover, bacteria treated with 0.4% EMS for 4 h and Etofibrate were significantly (p < 0.001) longer on average than unconstricted bacteria that were not exposed to EMS (Figure 3B). This suggests that growth is not arrested by the presence of EMS, while constriction is clearly inhibited. This is consistent with a replication arrest caused by alkylation of the bacterial genome, as previously reported for E. coli [22]. AidB polar localization persists inside host cells B. abortus is an intracellular pathogen that encounters various stresses during its life cycle [9]. Since these stresses could result in the alkylation of DNA, e.g. through nitrosative stress [14], we tested the localization pattern of AidB-YFP in B. abortus (XDB1120 strain) during an infection of human epithelial cells (HeLa cells).

The conditioned medium containing secreted SPARC protein suppressed the growth of pancreatic cancer cells, indicating that silencing of the SPARC gene may result in pancreatic

cancer development and progression [12]. In the current study, we detected the methylation levels and methylation pattern of the SPARC gene transcriptional regulation region (TRR) in normal, adjacent normal, chronic pancreatitis, and pancreatic cancer tissues to assess the altered methylation levels of the SPARC CB-5083 gene to determine if SPARC methylation can be used as a tumorigenesis marker for the early detection of pancreatic cancer. Methods Cell line and culture Pancreatic cancer cell line PANC1 was purchased from the American Type Culture Collection (Manassas, VA, USA) and PaTu8988 was a kind gift from Dr. H.P. Elsasser (Phillips University, Marburg, Germany). These cells were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (both were from Life Technologies Inc., Rockville, MD, USA) and incubated at 37°C in a humidified chamber with 95% air and 5% CO2. Patient tissue specimens A tissue and patient’s data usage protocol was approved by the Ethics Committee of our Selleck BAY 1895344 institution. Informed written consent was obtained from each patient. Tissue samples from 52 patients were obtained from the Second Military Medical University affiliated Changhai Hospital from August

2006 to December 2007; these samples were from 6 pathologically proven cases of chronic pancreatitis, PF-02341066 nmr 6 cases of normal pancreatic tissues, 40 cases of pancreatic cancer (ductal adenocarcinoma type), and corresponding normal tissue from those same 40 patients. The tissue samples were obtained and stored in liquid nitrogen

immediately after being resected in the operating room. For pancreatic cancer cases, tumor tissues that contained more than 70% tumor cells and the corresponding adjacent normal tissues without any tumor cell infiltration were selected. In addition, samples of white blood cells (WBCs) Olopatadine were obtained from two healthy volunteers. Clinicopathological data, including gender, age, status of tobacco smoking and alcohol consumption, tumor size, differentiation, lymph node metastasis, and TNM stages, were collected from the electronic medical records of the patients. Tobacco smoking was defined as at least one cigarette per day for no less than 1 year. Alcohol consumption was defined as intake of at least 50 ml of Chinese liquor, 250 ml of wine, or 500 of ml beer at least once a week for a minimum of 1 year. The 6th American Joint Committee on Cancer (AJCC) staging system was used to classify the clinical stage of pancreatic cancer. DNA extraction and bisulfite modification of DNA Genomic DNA from the tissues and cell lines was extracted using the phenol/chloroform method and precipitated with ethanol.

Lancet 2004, 364: 1757.CrossRefPubMed 85. Koutsky LA, Ault NSC 683864 purchase KA, Wheeler CM, Brown DR, Barr E, Alvarez FB, Chiacchierini LM, Jansen KU, Proof of Principle Study Investigators: A controlled trial of a human papillomavirus type 16 vaccine. N Engl J Med 2002, 347: 1645–1651.CrossRefPubMed 86. Miller AB, Hoogstraten B, Staquet M, Winkler A: Reporting results of cancer treatment. Cancer 1981, 47: 207–14.CrossRefPubMed 87. Therasse P, Arbuck SG, Eisenhauer EA, Wanders J, Kaplan RS, Rubinstein L, Verweij J, Van Glabbeke M, van Oosterom AT, Christian MC, Gwyther SG: New guidelines to evaluate the response to treatment

in solid tumours. J Natl Cancer Inst 2000, 92: 205–16.CrossRefPubMed 88. Rosenberg S, Yang JC, Restifo NP: Cancer immunotherapy: moving beyond current vaccines. Nat Med 2004, 10: 909–15.CrossRefPubMed 89. Nagorsen D, Thiel E: Clinical and immunologic responses to active specific cancer vaccines in human colorectal cancer. Clin Cancer Res 2006, 12: 3064–9.CrossRefPubMed 90. Mocellin S, Mandruzzato S, Bronte V, Marincola FM: Cancer vaccines: pessimism in check. Nat Med 2004, 10: 1278–80.CrossRefPubMed 91. Takeda K, Kaisho T, Akira

S: Toll-like receptors. Annu Rev Immunol 2003, 21: 335–376.CrossRefPubMed 92. Strome SE, Chen L: Costimulation-based immunotherapy for head and neck cancer. Curr Treat Options Oncol 2004, 5: 27–33.CrossRefPubMed 93. Rosenberg SA: Overcoming obstacles to the effective immunotherapy of human cancer. Proc check details Natl Acad Sci USA 2008, 105: 12643–4. Epub 2008 Aug; 27.CrossRefPubMed 94. Lappin M, Weiss J,

Delattre V, Mai B, Dittmar H, Maier C, Manke K, Grabbe S, Martin S, Simon JC: Analysis of mouse dendritic IMP dehydrogenase cell migration in vivo upon subcutaneous and intravenous injection. Immunology 1999, 98: 181–188.CrossRefPubMed 95. find more Kudo-Saito C, Schlom J, Hodge J: Induction of an antigen cascade by diversified subcutaneous/intratumoural vaccination is associated with antitumour responses. Clin Cancer Res 2005, 11: 2416–2426.CrossRefPubMed 96. Tagawa S, Lee P, Snively J, Boswell W, Ounpraseuth S, Lee S, Hickingbottom B, Smith J, Johnson D, Weber JS: Phase I study of intranodal delivery of a plasmid DNA vaccine for patients with stage IV melanoma. Cancer 2003, 98: 144–154.CrossRefPubMed 97. Bedrosian I, Mick R, Xu S, Nisenbaum H, Faries M, Zhang P, Cohen PA, Koski G, Czerniecki BJ: Intranodal administration of peptide-pulsed mature dendritic cell vaccines results in superior CD8+ T-cell function in melanoma patients. J Clin Oncol 2003, 21: 3826–3835.CrossRefPubMed 98. Williams T, Ynagimoto J, Mazumder A, Wiseman C: IL-2 increases the antibody response in patients receiving autologous intralymphatic tumour cell vaccine immunotherapy. Mol Biother 1992, 4: 66–69.PubMed Competing interests The author declares that they have no competing interests.

“
“Background Breast radiation therapy after conservative surgery is now widely accepted as a standard of care for patients with early GSK872 breast cancer. Moreover breast conserving therapy has become an accepted treatment option over radical mastectomy for stage I – II breast tumour [1–3]. The conventional radiation course consists of 50 Gy in 25 daily fractions of 2 Gy on the whole breast usually followed by the addition of a boost dose to the tumour bed of 10 to 16 Gy in 5 – 8 daily fractions resulting in overall 6 – 7 week treatment. However, in certain patient populations like the elderly and those living far from radiation facilities, adjuvant

breast radiotherapy appears to be underutilized because of the substantial length of treatment. Delivering postoperative radiotherapy in a shorter period of time could effectively be much more convenient for these patients.

That is, a shorter schedule of radiotherapy, as an accelerated GSK126 purchase hypofractionated regimen, could indeed improve the use of breast conserving therapy helping to knock down selleck chemical the “”logistical barriers”"(in terms of age, aged-related morbidity, time, travel difficulties, absence from family and job, cost etc) and consequently providing more women with this option. This accelerated hypofractionated approach is based on the radiobiologic model that a lower total dose delivered in fewer, larger fractions over a shorter period of time is at least as effective as the traditional longer schedule. The relationship between total dose, fraction size and tissue response is described by the α/β value (expressed in Gy) in Linear Quadratic (LQ) model [4]. Increasing evidence from randomized trials comparing conventional radiotherapy schedules

Tolmetin to different hypofractionated ones in whole breast irradiation after conserving surgery show that breast adenocarcinoma may be associated with lower α/β value than previously thought and closer to those of late-reacting healthy tissues [5–9]. The LQ model suggests that, when the α/β ratio for the tumour is similar to that of the surrounding late-responding normal tissue, the hypofractionated regimen may be equally or potentially more effective than the conventional one [10]. On this basis patients at our Institute who refused to spend 6 to 7 weeks in radiotherapy after breast conserving surgery were offered an accelerated hypofractionated radiation therapy schedule consisting of 10 daily fractions of 3.4 Gy to whole breast plus a boost dose of 8 Gy in a single fraction to the tumour bed. The paper aims to report a preliminary analysis focusing on the early and late skin and lung toxicity after this accelerated hypofractionated regimen. Lung toxicity was investigated in terms of CT density evaluation, pulmonary functional tests, and clinical and radiological scoring.

” This provision has led to a debate between WTO member states whether a revision of the WTO TRIPS Agreement is required to bring the agreement into line with the CBD in particular as far as the protection of traditional knowledge of local and indigenous communities mentioned in Article 8 (j) CBD is concerned. The TRIPS Agreement ��-Nicotinamide molecular weight does not make reference to traditional

knowledge. It does, however, require the granting of intellectual property rights to plant varieties, either in the form of patents or “by an effective sui generis system or by any combination thereof” (Article 27.3 (b) TRIPS). As for patents, the same provision of Article 27.3 (b) TRIPS allows for the exclusion from patentability of “plants and animals other than micro-organisms, and essentially biological processes for the production of plants or animals other than non-biological and micro-biological processes”. The provision aims at a fundamental distinction in patent law between non-patentable discoveries and inventions, which may be patented. The TRIPS selleck Agreement leaves it to national legislation where precisely to set the threshold

(Gervais 2003, p. 229). However, with the growth of the biotechnology industry, patenting of micro-organisms has become common following the decision of the US Supreme Court in Diamond v Chakrabarty (Rimmer 2008, pp. 24–49) and is now required in the TRIPS Agreement as is the patenting of non-biological and micro-biological processes. From its introduction, Article 27.3 (b) provided for a review of the provision four years after the Alectinib date

of entry into force of the WTO Agreement. While this mandate was reiterated at the Doha Ministerial Conference in 2001, the review has not generated any substantive results (Biber-Klemm et al. 2006, p. 79; Gervais 2003, pp. 227–234). In the international debate about the extension of intellectual property protection to plant varieties in particular, traditional knowledge has been used partly as a counterargument to defend regional, national and local interests especially related to food security and agriculture. It has further been used to raise counterclaims for the protection of knowledge more typically to be encountered in developing countries. The focus of this discussion has recently been on the proposal of a group of developing countries to require the disclosure in patent applications of the origin of any resources and/or associated knowledge used in generating an invention as well as evidence of prior informed consent and equitable benefit-sharing, a proposal which in turn triggered GSK2245840 alternative proposals from the US, Japan, the EU and Switzerland (Straus 2008, pp. 229–231). International definitions of “traditional knowledge” The precise definition of traditional knowledge is equally still debated.

1 months per patient; range 1 to +50.5. Patients treated in Switzerland were re-examined on average every other month for frequency detection; patients treated in Brazil were only examined once. Novel frequencies discovered upon re-examination were added to the treatment program of patients receiving experimental treatment. The first treatment programs consisted of combinations of less than ten frequencies while the most recent treatment programs exceed 280 frequencies (Figure 2). Figure 2 Compassionate treatment of a 51 year old patient with ovarian cancer FIGO IIIC with extensive

peritoneal carcinomatosis since October 1997. The patient received paclitaxel and cisplatin from March 97, then docetaxel and carboplatin, doxorubicin, and gemcitabine. Because of progression of disease the patient was offered compassionate treatment with amplitude-modulated KPT-8602 chemical structure electromagnetic TSA HDAC in vivo fields as of May 05. As seen below, the initial treatment consisting of 15 frequencies (May 05) did not yield any response. Upon re examination, 11 additional frequencies

(26) were added to the treatment program in August 05. Because of disease progression, treatment with single agent bevacizumab was initiated in November 05. Interestingly, the CA 125 level had decreased by 200 units prior to the PXD101 supplier initiation of bevacizumab. Combined treatment with amplitude-modulated electromagnetic fields and bevacizumab resulted in a decrease in CA 125 level from 2140 to 540 in May 06. Treatment was supplemented with cyclophosphamide from March to September 07. The patient was hospitalized with pneumonia and elected to only receive amplitude-modulated electromagnetic fields since September 07. As of April 09, i.e. 50.5 months after treatment initiation the patient has stable disease and is asymptomatic. The numbers above the arrows represent the total number of cancer-specific frequencies included in the treatment program. The evolution of treatment programs through incremental addition of tumor-specific frequencies is illustrated by the case of a 51 year old woman with ovarian cancer. This patient was diagnosed

with FIGO stage III (G2–G3) ovarian cancer in October 1997 and had received multiple courses of palliative chemotherapy until 2005. As seen on Figure 2, the initial treatment consisting of 15 frequencies did not yield any response. Upon re-examination, Tenofovir cost 11 additional frequencies (total of 26) were added to the treatment program in August 05. Because of disease progression, treatment with single agent bevacizumab was initiated in November 05. Interestingly, the CA 125 level had decreased by 200 units between October and November 2005, prior to the initiation of bevacizumab. Combined treatment with amplitude-modulated electromagnetic fields and bevacizumab resulted in a decrease in CA 125 level from 2140 to 540 in May 06. Treatment was supplemented with cyclophosphamide from March to September 07.

mobilis strains tested. However, their respective PCNs were closely matched within the same strain (Table 2). This indicated that the respective pUC18 or pACYC-184 derived plasmid backbones had little effect on their replication properties within Z. mobilis; which were primarily governed by the ca. 1,900 bp replicon fragment from pZMO7. Copy number was highest in the ATCC 29191 strain (ca. 20-40 plasmids per cell), and

considerably lower in the NCIMB 11163 and CU1 Rif2 strains (ca. 1-3 plasmids per cell). Further https://www.selleckchem.com/products/pu-h71.html detailed studies will be required to establish the physiological basis for this https://www.selleckchem.com/products/VX-680(MK-0457).html inter-strain variation in PCN. Protein expression and proteomic applications within Z. mobilis To demonstrate a proof of principle, we selected a well-established glutathione/glutathione S-transferase (GST) affinity ‘pull-down’ approach [34] for use in Z. mobilis. In addition

to functioning as a convenient method for one-step protein isolation, TSA HDAC supplier (N-terminal) GST fusions have previously been shown to be beneficial for the expression of soluble (heterologous) proteins in bacteria [48]. The AcpP, KdsA, DnaJ, Hfq and HolC proteins selected as ‘bait’ were included in a previous proteomic study conducted in E. coli[35]. With the exception of the Hfq RNA chaperone [49], the respective properties of these proteins have not previously been analyzed in Z. mobilis. Four out of five proteins were expressed in a soluble form in both the ATCC 29191 and CU1 Rif2 strains, clearly demonstrating the effectiveness of the pZ7-GST vector-based system. The GST-HolC protein may have been expressed in an insoluble form, thus failing to be recovered in the (soluble) cell lysate fractions. Co-purifying proteins ADP ribosylation factor were identified for two of the four GST-fusion proteins that were expressed

in the ATCC 29191 strain (AcpP and KdsA). However, it should be noted that the plasmid-based GST-fusion protein expression is performed in a wild-type chromosomal background. Consequently, the GST-tagged bait proteins will be in direct competition with the corresponding endogenous bait proteins, for the capture of binding partners within the cell. Hence it may not be possible to capture and purify sufficient levels of interacting protein species to enable their subsequent detection or identification. The acyl carrier protein AcpP, which acts as a covalent carrier of fatty acid intermediates during their biosynthesis, co-purified with four other functionally-related enzymes.

Plasmid pZM3H1 carries a large portion of this C. litoralis transposon (17 ORFs; orf8-orf23), DihydrotestosteroneDHT supplier although it lacks the 5.3 -kbterminal region of the element, which contains three genes coding for a putative NADP-specific glutamate dehydrogenase,

a conserved membrane protein and a transposase (Figure  1). This truncated transposon contains (i-ii) two heavy metal resistance cassettes – a Co/Zn/Cd efflux system (orf11, orf12) and mercury resistance determinants (orf16-orf22), (iii) an ORF encoding a protein of the metallo-beta-lactamase family (orf15), (iv) a site-specific resolution system (composed of two genes tnpS and tnpT, and a putative resolution site with a hairpin structure) homologous to the MRS system of Tn4651[51], as well as (v) four ORFs encoding hypothetical proteins with unknown functions (orf8, orf13, orf14 and orf23) (Figure  1). The putative efflux system (CZC ��-Nicotinamide module; orf11, orf12) encodes a predicted CzcD metal transport membrane protein (a member of the cation diffusion facilitator

protein family), which mediates cobalt (Co2+), zinc (Zn2+) and cadmium (Cd2+) resistance (as shown in Cupriavidus metallidurans CH34 [52]). The mercury resistance module (MER) contains 7 ORFs (orf16-orf22) with significant levels of homology to the merRTPABDE genes, responsible for enzymatic detoxification of Hg2+ ions to the less toxic form

Hg0[53]. The key enzymes in this mercury resistance system are (i) organomercurial lyase (MerB) – effectively performs hydrolysis of stable mercury-carbon bonds, and (ii) mercuric reductase (MerA) – reduces Hg2+ to Hg0 (metallic mercury) in a process that involves hydride transfer from the electron carrier NADPH to flavin. Three other important components are (i) two transcriptional regulatory proteins (MerR Smoothened and MerD), (ii) two mercury ion transport proteins (MerT and MerP), and (iii) an accessory membrane protein (MerE) [53] (Figure  1 and Additional file 1: Table S1). To investigate whether the analyzed resistance cassettes are functional, plasmid buy HM781-36B pBBR-ZM3CZCMER was constructed by inserting the orf11-orf23 gene cluster of pZM3H1 (contains the CZC and MER modules) into broad-host-range (BHR) mobilizable vector pBBR-MCS2 (see Methods for details). Since we were unable to remove (by incompatibility) plasmid pZM3H1 from its natural host (Halomonas sp. ZM3), the obtained plasmid pBBR-ZM3CZCMER was introduced (by conjugation or transformation) into Pseudomonas spp. LM7R and LM12R (pZM3H1 was shown to replicate in both strains) and E. coli TG1 (three members of Gammaproteobacteria), as well as A. tumefaciens LBA288 (Alphaproteobacteria).