阔褶水蛙皮肤组织cDNA文库的构建及抗菌肽brevinin-2LTs的cDNA克隆和序列分析

Construction of cDNA Library of Hylarana latouchii Skin Tissue with Cloning and Sequence Analysis of the cDNA Encoding Antimicrobial Peptides Brevinin-2LTs

运用SMART技术成功构建了阔褶水蛙（Hylarana latouchii）皮肤组织全长cDNA文库，用来筛选抗菌肽及其他功能性基因。经鉴定，原始文库滴度为9．7×10^6cfu／mL，重组率达到91％，插入片段大小在O．3～1．5kb范围内，文库扩增后的滴度为1．9×10^9cfu／mL，说明所构建文库质量较高。通过该文库成功筛选出2个抗菌肽cDNA序列，此序列属于brevinin-2家族，因此命名为brevirtin-2LT1和brevinin-2LT2。

A full length cDNA library of Hylarana latouchii skin tissue was constructed with the SMART technique. This cDNA library could be used to screen encoding gene of antimicrobial peptides and other functional genes. The titer of the cDNA library was 9.7 ×10^6 cfu/mL, and the recombination rate of it was 91%. Inserted fragments ranged was 0.3 - 1.5 kb. The titer of the amplified library was 1.9×10^9 cfu/mL.Successfully constructed cDNA library was a full length library with high quality. Two different cDNA sequences encoding antimicrobial peptide precursors named brevinin-2LT1 and brevinin-2LT2 were cloned by PCR from this cDNA library. All of the two antimicrobial peptides attributed to brevinin-2 family.