中华按蚊抗菌肽defensinA编码基因cDNA克隆与序列分析

目的克隆中华按蚊defensin编码基因,并对其序列进行分析。方法提取中华按蚊总RNA。据文献报道合成1对引物def1、def2,RT-PCR扩增中华按蚊defensin编码基因,克隆于T-载体,序列测定与分析。结果RT-PCR扩增中华按蚊cDNA获大约308 bp片段,测定目的片段序列,结果显示,克隆基因cDNA序列长度为308 bp,推导编码64个氨基酸,序列分析显示中华按蚊defensin编码基因与冈比亚按蚊defensin编码基因有高度同源性(95%)。分析结果显示,中华按蚊defensin编码基因为新基因,定名为中华按蚊defensinA。结论克隆出中华按蚊defensinA编码基因,为进一步研究该基因打下了基础。

CLONING AND SEQUENCE ANALYSIS OF cDNA ENCODING THE ANTIBACTERIAL PEPTIDE, DEFENSINA FROM THE MOSQUITO, ANOPHELES SINENSIS

Objective To clone cDNA encoding the antibacterial peptide, defensinA from mosquito Anopheles sinensis and conduct sequence analysis.Methods Total RNA was extracted from An.sinensis,primers def1 and def2 were synthesized according to literary,RNA sequence of the adult of An.sinensis was amplified by RT-PCR to get the defensin cDNA which was then cloned into T vector and sequenced.The cDNA sequence of defensin encoding gene was analysed and compared with other defensin gene of insect.Results It was found that a DNA fragment encoding defensin about 308 bp in length was obstained from An.sinensis cDNA.Its 308 bp nucleotide sequences and deduced 64 amino sequences shared highly identity with An.gambiae defensin encoding sequence(95%).It was evaluated An.sinensis defensin encoding gene is a new gene.It was given a name is An.sinensis defensisA.Conclusion An.sinensis defensinA encoding gene have been successfully cloned.These will provide for a basis to study the gene.