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Findings from Biotechnology Research Institute in the Area of Plasmid Research Reported (Construction of a standard reference plasmid containing...

August 29, 2014



Findings from Biotechnology Research Institute in the Area of Plasmid Research Reported (Construction of a standard reference plasmid containing seven target genes for the detection of transgenic cotton)

By a News Reporter-Staff News Editor at Science Letter -- Fresh data on Plasmid Research are presented in a new report. According to news reporting from Beijing, People's Republic of China, by NewsRx journalists, research stated, "Insect resistance and herbicide tolerance are the dominant traits of commercialized transgenic cotton. In this study, we constructed a general standard reference plasmid for transgenic cotton detection."

The news correspondents obtained a quote from the research from Biotechnology Research Institute, "Target genes, including the cowpea trypsin gene cptl, the insect resistance gene cry1Ab/1Ac, the herbicide tolerance gene c714-epsps, the Agrobacterium tumefaciens nopaline synthase (Nos) terminator that exists in transgenic cotton and part of the endogenous cotton Sadl gene were amplified from plasmids pCPT1, pBT, pCP4 and pBI121 and from DNA of the nontransgenic cotton line K312, respectively. The genes crylAbIlAc and cpti, as well as cp4-epsps and the Nos terminator gene, were ligated together to form the fusion genes cpti-Bt and cp4-Nos, respectively, by overlapping PCR. We checked the validity of genes Sadl, cpti-Bt and cp4-Nos by DNA sequencing. Then, positive clones of cptl-Bt, cp4-Nos and Sadl were digested with the corresponding restriction enzymes and ligated sequentially into vector pCamBIA2300, which contains the CAMV 35S promoter and nptil gene, to form the reference plasmid pMCS. Qualitative detection showed that pMCS is a good positive control for transgenic cotton detection. Real-time PCR detection efficiencies with pMCS as a calibrator ranged from 9435% to 98.67% for the standard curves of the target genes (R2 0.998). The relative standard deviation of the mean value for the known sample was 11.95%. These results indicate that the strategy of using the pMCS plasmid as a reference material is feasible and reliable for the detection of transgenic cotton."

According to the news reporters, the research concluded: "Therefore, this plasmid can serve as a useful reference tool for qualitative and quantitative detection of single or stacked trait transgenic cotton, thus paving the way for the identification of various products containing components of transgenic cotton."

For more information on this research see: Construction of a standard reference plasmid containing seven target genes for the detection of transgenic cotton. Plasmid, 2014;74():39-44. Plasmid can be contacted at: Academic Press Inc Elsevier Science, 525 B St, Ste 1900, San Diego, CA 92101-4495, USA. (Elsevier - www.elsevier.com; Plasmid - www.elsevier.com/wps/product/cws_home/622933)

Our news journalists report that additional information may be obtained by contacting X.J. Wang, Chinese Academy Agr Sci, Biotechnol Res Inst, Beijing 100081, People's Republic of China. Additional authors for this research include Q.L. Tang, L. Dong, Y.F. Dong, Y.Y. Su, S.R. Jia and Z.X. Wang (see also Plasmid Research).

Keywords for this news article include: Asia, Beijing, Plasmid Research, People's Republic of China

Our reports deliver fact-based news of research and discoveries from around the world. Copyright 2014, NewsRx LLC


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Source: Science Letter


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