中国农业科学院农业信息研究所 中国农学会
Agricultural Information Institute of CAAS China Association of Agricultural Science Societies
En
Ch
中国农业科学院农业信息研究所 中国农学会 主办

AI检索

订阅

返回顶部
当前位置: 首页 > 文章 > 家蝇抗菌肽天蚕素基因的定点突变和高效表达质粒的构建 蚕业科学 2004,30 (1) 92-96
Position: Home > Articles > The Site-directed Mutagenesis and Construction of A Highly Productive Expression Vector of the Cecropin Gene, An Antimicrobial Peptide From the Housefly (Musca domestica) Science of Sericulture 2004,30 (1) 92-96

家蝇抗菌肽天蚕素基因的定点突变和高效表达质粒的构建

作  者：
许小霞;徐兴耀;金丰良;张古忍;张文庆
单  位：
中山大学生物防治国家重点实验室;华南农业大学动物科学学院
关键词：
家蝇;天蚕素;反转录聚合酶链式反应;开放阅读框;pPICZαA
摘  要：
运用反转录聚合酶链式反应 (RT PCR)技术从家蝇体内扩增出抗菌肽天蚕素 (cecropin)基因的开放阅读框(ORF) ,与pMD 18T载体重组 ,经限制性酶切片段分析和核苷酸序列分析 ,与GenBank中报道的序列一致。根据此ORF ,重新合成 1对引物 ,并在碳末端进行定点突变 ,加上Asn编码 ,使其末端酰氨化 ,再利用半嵌套式PCR扩增出家蝇cecropin基因的成熟肽 ,与双酶切的酵母表达载体pPICZαA连接 ,经PCR和双酶切鉴定 ,成功构建了分泌型表达质粒。
译  名：
The Site-directed Mutagenesis and Construction of A Highly Productive Expression Vector of the Cecropin Gene, An Antimicrobial Peptide From the Housefly (Musca domestica)
作  者：
XU Xiaoxia 1,2 XU Xingyao 2 JIN Fengliang 1 ZHANG Guren 1 ZHANG Wenqing 1( 1State Key Lab for Biocontrol, Sun Yat-sen University, Guangzhou 510275; 2College of Animal Science, South Agricultural University, Guangzhou 510642)
关键词：
Housefly (Musca domestica)CecropinRT-PCRORFpPICZαA
摘  要：
The total RNAs were isolated from the larvage of houseflies (Musca domestica). The open reading frame (ORF) of Cecropin gene were cloned by using reverse transcriptase polymerse chain reaction (RT-PCR) from larvae of houseflies (Musca domestica). The PCR products were ligated into pMD-18T vector. According to biological softwares and the ORF of cecropin, the mature peptide were cloned by using RT-PCR, Nested PCR methods. The nucleotide sequence of the fragment, mature cecropin, which consists of 111 bp , encods 37 amino acid residues. The PCR products were ligated into pMD-18T vector too. The positive recombinant were digested by XhoⅠ/NotⅠ, the digested PCR products can be ligated into pPICZαA vector which were also digested by XhoⅠ/NotⅠ, then were transformed into DH5 α. It is indicated that we have succeeded constructing a productive expression vector, pPICZαA-cecropin.

相似文章

导出引用
计量
文章访问数: 9
HTML全文浏览量: 0
PDF下载量: 0

所属期刊

推荐期刊

友情链接Friendly link
农业信息研究所Agricultural information institute 农业智能知识服务平台Agricultural intelligent knowledge service platform 农业学术服务平台Agricultural Knowledge Service System 中国知网CNKI 维普网VIP 万方WANFANG DATA
联系我们Contact us

电话Tell：010-82109652-808

邮箱E-mail：agrijournal@caas.cn

服务Service
关于我们About us 意见建议Opinions and suggest
版权说明Copyright
单位：中国农业科学院农业信息研究所 中国农学会 © 2017中国农业科学院农业信息研究所 ICP备案号：京ICP备09089781号-46