Construction and Characterization of a Fosmid Library of Vibrio cholera Serogroup O1
A fosmid library was constructed and characterized for Vibrio cholera serogroup Ol in this study. Intact genomic DNA of Vibrio cholera N16961 was extracted with low melting point agarose embedment and β-agarase I digestion methods, fragmented with pipette and separated through pulse field gel electrophoresis. The fragments with desirable lengths ranging from 33 to 48kb were recovered from agarose gel with the ends blunted and phosphorylated. These fragments were ligated with pCClFOS vector, packaged and and transfected into hosting bacteria, yielding a fosmid library containing 9600 clones. The inserts were 37kb in length in average. Vector containing clones were less than 2%. The constructed reached the standard of a fosmid library, which will certainly facilitate cloning the genes of Vibrio cholera N16961 and deciphering their functions.
国家科技期刊平台
NSTL
万方数据
维普
