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 > 鹅生长激素基因的克隆、原核表达及其重组蛋白质的离子交换纯化 江苏农业学报 2014 (6) 1387-1391
Position: Home > Articles > Cloning and prokaryotic expression of goose growth hormone gene and recombinant protein anion-exchange purification Jiangsu Journal of Agricultural Sciences 2014 (6) 1387-1391

鹅生长激素基因的克隆、原核表达及其重组蛋白质的离子交换纯化

作  者：
李辉;陈蓉;应诗家;施振旦;赵伟
单  位：
江苏省农业科学院畜牧研究所
关键词：
鹅;生长激素;原核表达;阴离子交换
摘  要：
为了克隆鹅生长激素基因并表达其重组蛋白质,采集生长期鹅垂体组织,并利用TRIzol快速提取的总RNA为模板,反转录为c DNA。根据鹅生长激素基因编码的成熟肽序列(Gen Bank号:AY149895.2)设计1对引物,分别在上、下游引物的5'端引入Nhe I和Hind III酶切位点。经反转录扩增获得鹅生长激素基因的编码的成熟肽全序列。通过双酶切和连接将鹅生长激素编码区插入原核表达载体pRSET-A的Nhe I和Hind III位点之间,构建重组表达质粒pRSET-g GH并转化大肠杆菌表达菌株BL21(DE3)。转化的菌株经IPTG诱导后表达重组鹅生长激素蛋白质,分子量约为2.93×104。经过DEAE-650M弱阴离子交换树脂纯化获得较高纯度的重组鹅生长激素蛋白质。
译  名：
Cloning and prokaryotic expression of goose growth hormone gene and recombinant protein anion-exchange purification
作  者：
LI Hui;CHEN Rong;YING Shi-jia;SHI Zhen-dan;ZHAO Wei;Institute of Animal Science,Jiangsu Academy of Agricultural Sciences;
关键词：
goose;;growth hormone;;prokaryotic expression;;anionexchange
摘  要：
Total RNA of goose growth hormone gene extracted from goose pituitary tissue with TRIzol reagent were used as template for reverse transcription of the first strand c DNA and a pair of primers was designed based on the published goose growth hormone gene sequence fragment(Gen Bank No.AY149895.2),in which,two restriction enzyme sites Nhe I and Hind III were introduced into the 5' end of both upstream and downstream primers respectively.Goose growth hormone gene coding sequence was amplified by PCR,and the amplified mature peptide sequence was inserted into the Nhe I and Hind III sites of the expression vector pRSET-A to generate the recombinant expression plasmid pRSET-g GH which was transformed into Escherichia coli BL21(DE3) afterwards.The transformed bacterium was induced with IPTG to express the recombinant protein with a molecular mass of 2.93×104.The high purity recombinant goose growth hormone was achieved using DEAE-650 M weak anion exchange resin.

相似文章

导出引用
计量
文章访问数: 21
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