%0 Journal Article
%T Construction of double-labelled carbofuran-degrading bacterium Sphingomonas sp. CDS-1<br>呋喃丹降解菌CDS-1的双标记菌株的构建
%A XU Jian-hong
%A WU Jun
%A HONG Qing
%A ZHANG Zhi-lin
%A LI Shun-peng
%A WANG Yun-duan
%A <br>徐剑宏
%A 武俊
%A 洪青
%A 张志琳
%A 李顺鹏
%J 微生物学报
%D 2006
%I 
%X The genomic DNA of a carbofuran-degrading bacterium Sphingomonas sp. CDS-1 was digested by Sau3Al and ligated to pRobe-GFP digested by BamHI, and the product was transformed to the E. coli DH5alpha competent cells. Fifty positive clones that could emit green fluorescence under UV were selected from about 1 x 10(4) clones grown on selective plates AmpLB. One clone F7 with the strongest fluorescence was selected, the recombinant plasmid pF7 from this clone was digested with EcoR I & Hind III and the DNA fragment comprising gfp and promoter of Sphingomonas sp. CDS-1 was recovered, which was subsequently cloned into the broad host vector pPZP201 to construct a new plasmid pPZP201-gfp. pPZP201-gfp was introduced into Sphingomonas sp. strain CDS-1 by triparental conjugation to make strain CDS-gfp. gfp was expressed strongly and stably in strain CDS-gfp after 10 times successive re-culturing (48 h/time). The linA gene was inserted into Not I -cut transposon vector pUT/mini-Tn5 to construct a new transposon vector pUT/mini-Tn5-linA. With the aid of helper plasmid pRK600, pUT/mini-Tn5-linA was introduced into CDS-gfp, the dehydrochlorinase gene linA was integrated into the chromosome of CDS-gfp by transposing. The double labelled strain CDS-GFP-LinA was constructed. This strain was also a genetic engineering strain that was able to degrade gamma-hexachlorocyclohexane and carbofuran simultaneously. All of these results laid a foundation for the study of ecological performance of Sphingomonas sp. CDS-1.
%K Sphingomonas sp
%K CDS-1
%K Carbofuran
%K Degradation
%K Double-labelled
%K Genetic engineering strain<br>Sphingomonas
%K sp.CDS-1
%K 呋喃丹
%K 降解
%K 双标记
%K 基因工程菌
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=D87A9304FCA1712D&yid=37904DC365DD7266&vid=D997634CFE9B6321&iid=E158A972A605785F&sid=9E7C0CB25117E09B&eid=7FAAB0292FA0D5D0&journal_id=0001-6209&journal_name=微生物学报&referenced_num=1&reference_num=16