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小尾寒羊YB-1基因全长cDNA克隆与原核表达 被引量：1

Cloning of Small-tailed Han Sheep YB-1 cDNA and Expression in E.coli
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摘要 采用RT-PCR技术从小尾寒羊睾丸组织获得YB-1基因全长cDNA,并将其编码区重组于融合表达质粒pET32a中,经酶切和序列鉴定分析后,重组质粒在大肠杆菌BL21 plys(E)中经不同浓度IPTG诱导后获得表达,但表达量较低,且IPTG的浓度变化不影响目的蛋白的表达量。通过改变筛选抗生素,最终获得了目的融合蛋白的高效表达,为深入开展YB-1蛋白功能研究打下了良好的基础。 Full-length cDNA of YB-1 gene was cloned from testicle in Small-tailed Han sheep by RT-PCR. pET32a expression system was used to express open reading frame （ORF） of YB-1 in E. coli with induction of IPTG at different concentrations. The low expression level of fusion pro- tein revealed that IPTG concentration did not affect on its expression level. To improve the expression level of fusion protein, ampicillin was replaced by carbenicillin in cultural medium, which increased expression efficiency.
作者 宋雪梅 李宏滨 杜立新
机构地区 中国农业科学院北京畜牧兽医研究所
出处 《畜牧兽医学报》 CAS CSCD 北大核心 2008年第3期365-367,共3页 ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金 "十一五"国家科技支撑计划(2006BAD01A11) 国家"863"计划(2006AA10Z199)
关键词 小尾寒羊 YB-1 融合蛋白 原核表达 Small-tailed Han sheep YB-1 fusion protein prokaryotic expression
分类号 S828.2 [农业科学—畜牧学]
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同被引文献20

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畜牧兽医学报
2008年 第3期

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