新疆細(xì)毛羊eIF3l基因的克
發(fā)布時(shí)間:2018-03-02 12:18
本文選題:新疆細(xì)毛羊 切入點(diǎn):eIFl基因 出處:《畜牧與獸醫(yī)》2017年08期 論文類型:期刊論文
【摘要】:為了探究新疆細(xì)毛羊真核生物翻譯起始因子(eukaryotic translation initiation factors,eIFs)基因eIF3l序列和原核表達(dá)蛋白特征,采用RTPCR法從新疆細(xì)毛羊成纖維細(xì)胞中克隆eIF3l基因mRNA的CDS區(qū)編碼序列,構(gòu)建其原核表達(dá)載體,并進(jìn)行原核表達(dá)和蛋白檢測(cè)。結(jié)果顯示:完整獲得eIF3l基因1 695 bp,共編碼564個(gè)氨基酸殘基;37℃,1 mmol/L異丙基硫代半乳糖苷(IPTG)誘導(dǎo)3.5 h可形成大量包涵體蛋白,其分子量約為69 ku;Western blot鑒定目的蛋白表達(dá)正確。結(jié)果表明:新疆細(xì)毛羊eIF3l基因原核表達(dá)成功,為進(jìn)一步研究其功能和應(yīng)用提供了科學(xué)數(shù)據(jù)。
[Abstract]:In order to investigate the eIF3l sequence of eukaryotic translation initiation factors (IFS) gene and prokaryotic expression protein in Xinjiang fine wool sheep, the CDS coding sequence of eIF3l gene mRNA was cloned from fibroblasts of Xinjiang fine wool sheep by RTPCR method. The prokaryotic expression vector was constructed, and the prokaryotic expression and protein detection were performed. The results showed that 1 695 BP of eIF3l gene encoding 564 amino acid residues at 37 鈩,
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