本文已被：浏览 6017次 下载 624次
投稿时间：2021-03-10
投稿时间：2021-03-10
中文摘要: 以谷氨酸棒杆菌Corynebacterium glutamicum ATCC 13032基因组为模板，通过聚合酶链式反应（polymerase chain reaction，PCR）扩增得到DNA分子量大小为1 815 bp和1 700 bp的乙酰辅酶A羧化酶α-亚基以及β-亚基，将目的基因连接到pXMJ19质粒，得到表达载体pXMJ19-accBC、pXMJ19-accD1，将其转化至表达菌株BL21，得到异源表达菌株BL21-accBC、BL21-accD1。使用气相色谱测定发酵液中脂肪酸含量。结果显示：重组菌株BL21-accBC与BL21-accD1经1 mmol/L的异丙基-β-D-硫代半乳糖苷有氧诱导20 h后，脂肪酸含量分别达到45.57 mg/g菌体干重和18.81 mg/g菌体干重，较对照菌株分别提高了266%和51.32%。
Abstract:In this study，the genome of Corynebacterium glutamicum ATCC 13032 was used as a template，and coding sequences for acetyl-CoA carboxylase α-and β-subunits （1 815 bp and 1 700 bp，respectively）were obtained by polymerase chain reaction（PCR）amplification.Target genes were ligated into the expression vector pXMJ19 to obtain expression vectors pXMJ19-accBC and pXMJ19-accD1.These vectors were transformed into the E.coli expression strain BL21 to obtain heterologous expression strains BL21-accBC and BL21-accD1.The fatty acid content in the fermentation broth was determined by gas chromatography.The results showed that the fatty acid output of the recombinant strains BL21-accBC and BL21-accD1 reached 45.57 mg/g dry cell weight and 18.81 mg/g dry cell weight after aerobic induction with 1 mmol/L isopropyl-β-D-thiogalactoside（IPTG）for 20 hours.Compared with the control strain，fatty acids in recombinant Escherichia coli BL21-accBC and BL21-accD1 increased by 266%and 51.32%respectively.
keywords: Corynebacterium glutamicum acetyl-CoA carboxylase fatty acid Escherichia coli heterologous expression
文章编号：202123024 中图分类号： 文献标志码：
基金项目:国家自然科学基金（31860439）
| Author Name | Affiliation |
| YANG Qing，SUN Zi-yu，Mandlaa，WANG Jia，LIU Jin，JIN Jing-jing，CHEN Zhong-jun | Inner Mongolia Agricultural University，Hohhot 010018，Inner Mongolia，China |
引用文本：
津ICP备20000603号-1