ApCl基因在毕赤酵母中的表达  

Inducible Expressing of ApCl Gene in Pichia pastoris

作　　者：江程记[1,2] 高建明[2] 刘巧莲[2] 代真真[1,2] 郑金龙[2] 易克贤[2,3] 

机构地区：[1]海南大学农学院,海口571101 [2]中国热带农业科学院热带生物技术研究所农业部热带作物生物技术重点开放实验室,海口571101 [3]中国热带农业科学院环境与植物保护研究所,儋州571737

出　　处：《生物技术通报》2011年第9期86-89,共4页Biotechnology Bulletin

基　　金：农业部"948"项目(2010-Z6);中央级公益性科研院所基本科研业务费(ITBBZD0828);国家科技支撑计划(2007BAD48B06-05);海南省自然科学基金项目(80510);国家麻类现代农业技术体系建设专项(nycytx-19)

摘　　要：为了研究ApCl基因的功能,应用基因重组技术将ApCl基因片段克隆入大肠埃希菌―酵母穿梭质粒pGAPZαA,构建重组真核表达质粒pGAPZαA-ApCl,电转化巴斯德毕赤酵母GS115,Zeocin筛选出阳性克隆转化子。通过比较转化空载体pGAPZαA和重组载体pGAPZαA-ApCl的不同菌株分别在含有高盐和高山梨醇浓度的液体培养基中生长情况,发现毕赤酵母GS115在转化ApCl基因后其抗旱、抗盐能力显著提高(约3倍),进一步验证了ApCl基因对提高生物抗逆能力有显著作用,为将来分离该蛋白及进一步研究奠定基础。For research ApCl gene function,the DNA ApCl gene sequence was subcloned into yeast shuttle expression vector pGAPZαA,and the recombinant plasmid pGAPZαA-ApCl was constructed.Pichia pastoris GS115 was electrotransformed with pGAPZαA and pGAPZαA-ApCl,and the postive transformants were selected in the medium of zeocin.Then,the two electrotransformed yeasts with different vectors were cultured in two kinds of mediums that have high levels of salt and sorbitol.Compared to Pichia pastoris GS115 with pGAPZαA,transgenic yeast with ApCl gene grows faster,about there times of the contrast.It is proved that ApCl gene is significant to prove biology anti-adversity.The finding settles a favourable base to study and research ApCl protein.

关 键 词：ApCl基因 抗旱 抗盐 巴斯德毕赤酵母 生物抗逆 

分 类 号：Q78[生物学—分子生物学]