大腹圆蛛主壶腹腺cDNA文库构建和丝蛋白基因筛选

Construction of araneus ventricosus major ampullate gland cDNA library and screening spider silk gene

首次通过反转录 -置换法和使用pUC18质粒成功构建大腹圆蛛 (Araneusventricosus)主壶腹腺 (majorampullategland)cDNA基因文库 ,并以鸟枪法从中筛选出具有典型重复结构的大腹圆蛛主壶腹丝蛋白cDNA基因AvF1,大小为 174 4bp ,编码区为15 72bp ,编码氨基酸 5 2 4个 ,分子量为 4 2 4 89 5 5Da,典型的重复结构为(GGP)nGGX。与现有已知的蛛丝蛋白基因中三带金蛛 (Argiopetrifas ciata)鞭毛样丝基因 (AtfF)有最高的同源性 6 9 3%。大腹圆蛛主壶腹腺cDNA文库的构建和蛛丝蛋白新基因的克隆 ,为提供大腹圆蛛蛛丝蛋白基因背景和进一步研究蛛丝蛋白奠定了基础。

The cDNA library of major ampullate of Araneus ventricousus was prepared successfully for the first time by using pUC18 vector in the world. The gene(AvF1) encoding spider silk protein was cloned by means of picking colony randomly (bird gun). AvF1 is 1744bp and its region in encoding is 1572bp. Protein encoded by AvF1 is 524 amino-acids and molecular weight is 42489.55dal. Its repetitive motif typically is (GGP)nGGX and the homology with Argiope trifasciata flagelliform silk protein mRNA(AtfF) is 69.3%, which is the maximum comparing with all silk genes known in NCBI GeneBank. Construction of Araneus ventricousus ampullate gland cDNA library and the finds of the new spider silk genes are the bases of supplying the background of Araneus ventricousus silk genes and further researching in the spider silk proteins.