黄粉虫抗冻蛋白基因TmAFP84的克隆和序列分析

Cloning and Sequence Analysis of Antifreeze Protein Gene TmAFP84 in the Beetle Tenebrio molitor

对黄粉虫抗冻蛋白(AFPs)基因进行RT-PCR扩增后,构建克隆重组质粒pUCm-T-TmAFP84,经双酶切、PCR和测序鉴定,克隆出了与GenBank中公布的编码84个氨基酸基因(注册号AF159114)基本相一致的抗冻蛋白基因TmAFP84,其同源性高达98.81%.与其他编码84个氨基酸的抗冻蛋白基因的同源性为95.92%.进而构建了表达重组质粒pMAL-p2X-TmAFP84.

Tenebrio molitor antifreeze protein gene TmAFP84 was obtained through RT-PCR method. Then we constructed cloning recombinant plasmid pUCm-T-TmAFP84, which was identified by double restriction enzymes digestion analysis, PCR and sequencing. The sequence of TmAFP84 was compared with relative nucleotides reported in GenBank. Display： TmAFP84 shared high homology with other AFP gene of Tenebrio molitor, the value is up to 98.81%. We also constructed expressing recombinant vector pMAL-p2X-TmAFP84 for the future protein expression and research on the structure and the function.