ABSTRACT
Beijing has been one of the epicenters attacked most severely by the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) since the first patient was diagnosed in one of the city's hospitals. We now report complete genome sequences of the BJ Group, including four isolates (Isolates BJ01, BJ02, BJ03, and BJ04) of the SARS-CoV. It is remarkable that all members of the BJ Group share a common haplotype, consisting of seven loci that differentiate the group from other isolates published to date. Among 42 substitutions uniquely identified from the BJ group, 32 are non-synonymous changes at the amino acid level. Rooted phylogenetic trees, proposed on the basis of haplotypes and other sequence variations of SARS-CoV isolates from Canada, USA, Singapore, and China, gave rise to different paradigms but positioned the BJ Group, together with the newly discovered GD01 (GD-Ins29) in the same clade, followed by the H-U Group (from Hong Kong to USA) and the H-T Group (from Hong Kong to Toronto), leaving the SP Group (Singapore) more distant. This result appears to suggest a possible transmission path from Guangdong to Beijing/Hong Kong, then to other countries and regions.
Subject(s)
Humans , Genome, Viral , Haplotypes , Mutation , Open Reading Frames , Phylogeny , Severe acute respiratory syndrome-related coronavirus , GeneticsABSTRACT
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.