冬鲽鱼抗冻蛋白基因的人工合成  

Artificial synthesis of winter flounder antifreeze protein gene

作　　者：黄璐[1] 黄乐平 袁松华[1] 白利雄[1] 李新平[1] 

机构地区：[1]苏州大学生命科学学院,江苏苏州215123 [2]新疆乌鲁木齐农科院核技术生物技术研究所,新疆乌鲁木齐830000

出　　处：《苏州大学学报（自然科学版）》2007年第3期80-83,共4页Journal of Soochow University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30471250)

摘　　要：合成适合在棉花中表达的冬鲽鱼抗冻蛋白(Winter flounder antifreeze protein,Wf-AFP)基因.参照冬鲽鱼抗冻蛋白的氨基酸序列及棉花偏爱的密码子,重新设计适合在棉花中表达的Wf-AFP基因序列;根据此序列设计6条寡核苷酸引物,通过PCR方法将其拼接成完整的基因;克隆到pUCm-T载体上,转化大肠杆菌DH5α,抽提重组子进行酶切鉴定及序列分析.结果表明合成的Wf-AFP基因序列与设计方案完全相符,通过PCR介导的基因合成方法成功获得了适合在棉花中表达的冬鲽鱼抗冻蛋白基因.To synthesize winter flounder antifreeze protein（Wf-AFP） gene.Wf-AFP cDNA was redesigned according to its amino acid sequence and the high-usage condons in cotton.Six overlapping oligonucleitides fragments were designed and synthesized,and then assembled by recursive PCR.The synthesized gene was inserted into pUCm-T plasmid,which was transferred into the competent cell E.coil DH5α,and positive recombinants were screened.Sequence analysis showed that the synthesized gene was identical with the designed gene sequence.The result showed that we had successfully obtained the whole gene of Wf-AFP,which will provide material for high-level of expression in cotton.

关 键 词：冬鲽鱼抗冻蛋白 基因合成 PCR 

分 类 号：Q78[生物学—分子生物学]