人骨形态发生蛋白4多拷贝表达盒的构建及其表达  

Construction and Expression of Multiple-copy Expression Cassette of Human Bone Morphogenetic Protein 4

作　　者：黄义德[1,2] 吴金芸[1] 徐轶彦[3] 韩明[1] 林政[1,2] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108 [2]福建省发育与神经生物学重点实验室,福建福州350108 [3]福建卫生职业技术学院,福建福州350101

出　　处：《福建师范大学学报（自然科学版）》2011年第6期66-70,共5页Journal of Fujian Normal University：Natural Science Edition

基　　金：福建省自然科学基金资助项目(2009J01127);福建省教育厅资助项目(JB09038)

摘　　要：为了了解在毕赤酵母中人骨形态发生蛋白4基因剂量对其表达量的影响,利用毕赤酵母表达载体pAO815质粒上的2个同尾酶BglⅡ和BamHⅠ在体外构建了人骨形态发生蛋白4成熟肽序列1,3,6,9个拷贝表达盒,并对它们进行表达量研究.结果表明,在毕赤酵母中表达人骨形态发生蛋白4蛋白时,1个拷贝基因能达到最大表达量,增加人骨形态发生蛋白4基因的拷贝数并不会增加人骨形态发生蛋白4的表达量,基因拷贝数的增加反而导致人骨形态发生蛋白4在毕赤酵母中的表达水平下降.To understand the effect of gene dosage of human bone morphogenetic protein 4（hBMP4） on expression levels in Pichia pastoris,recombinant plasmids carrying 1,3,6,9 expression cassette copies of hBMP4 mature peptide DNA sequence were constructed with unique Bgl Ⅱ and BamH Ⅰ sites shared 4 bases in common between their recognition sites in pAO815 vector in vitro.Then these recombinant plasmids were transformed into Pichia pastoris to analyze expression levels of hBMP4 in Pichia pastoris.The results showed that the highest expression levels of recombinant hBMP4 were achieved by a 1-copy strain,expression levels of hBMP4 were found to decrease for high copy strains in Pichia pastotis.

关 键 词：毕赤酵母 人骨形态发生蛋白4 基因拷贝数 

分 类 号：R394.2[医药卫生—医学遗传学]