Cloning and characterization of the 16s rRNA of six species in the bacteria related with the infection of respiratory tract / 中国免疫学杂志

ABSTRACT

Objective:To clone and characterize the 16S rRNA of six species in the bacteria infecting respiratory tract to make gene chip.Methods:The primers of the target gene were designed and synthesized,and then the aimed fragment of the 16s rRNA was amplified by PCR and cloned.Finally the recombinant plasmids were characterized.Results:(1)The 16s rRNA gene of six species of bacteria was amplified.It was found that the size of amplified product by PCR was 1 300 bp in E.coli,S.aureus,S.pneumoniae,K.pneumoniae and H.influenzae and that of 1 100 bp in P.aeruginosa.(2)The JM109 transferred by the recombinant plasmid pMD18-T grew in Ampr culture was white colonies.(3)The specific bands could be found by restriction endonuclease and PCR analysis. (4)The sequence of the six bacterial 16s rRNA showed the same as those in the GenBank.Conclusion:The 16s rRNA of six species of bacteria is successfully amplified and cloned into plasmid pMD18-T. It will provide the basis for making gene chip detecting the six species of bacteria infecting respiratory tract.