[Cloning and sequence analysis of SSU rRNA gene of cutaneous leishmaniasis pathogen from Xinjiang of China].

OBJECTIVE

By sequencing of SSU rRNA gene cloning from Xinjiang cutaneous leishmaniasis pathogen (XJCLP) to provide evidence for identification of the pathogen.

METHODS

By PCR assay with primers R222 and R333, the specific fragment had been produced from SSU rRNA gene of XJCLP, L. infantum, L. tropica and cloned into pGEM-T Easy vector. The clones were sequenced by the Sanger dideoxy-mediated chain termination method, analysis of SSU rRNA gene sequences from XJCLP, L. tropica, L. infantum with DNASIS.

RESULTS

Sequence analysis showed that the specific fragment of SSU rRNA gene from XJCLP, L. infantum, L. tropica, were all 394 bp in length. There were 391 bases identical and three point mutations between the sequences of XJCLP and L. tropica, the similarity being 99.2%; 390 bases identical and three point mutations and one insertion/deletion between the sequences of XJCLP and L. infantum, the similarity being 99.0%. One insertion/deletion between the sequences of L. tropica and L. infantum, the similarity being 99.7%. The primary and secondary structures of SSU rRNA gene from XJCLP differed from those of L. infantum and L. tropica. A retrieval from GenBank confirmed that these 394 bp sequence are new gene sequences.

CONCLUSION

The primary and secondary structures of SSU rRNA gene from XJCLP, L. infantum, L. tropica were different. 394 bp sequence from SSU rRNA gene of XJCLP is a new gene sequence.