谷氨酸棒杆菌高丝氨酸脱氢酶编码基因hom的敲除

中国生物工程杂志

2007, Vol. 27

Issue (1): 59-63

研究报告

谷氨酸棒杆菌高丝氨酸脱氢酶编码基因hom的敲除

饶志明¹,张君胜²,沈微³,方慧英²,诸葛健³

1. 江南大学工业生物技术教育部重点实验室、江南大学生物工程学院工业微生物研究中心
2. 江南大学工业微生物研究中心
3.

Disruption of hom gene encoding for homoserine dehydrogenase of Corynebacterium glutamicum

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摘要： 本研究以谷氨酸棒杆菌（Corynebacterium glutamicum）标准菌株ATCC 13032染色体为模板，设计引物PCR扩增高丝氨酸脱氢酶编码基因（hom），在hom基因内部插入一段来源于质粒pET28a的卡那霉素抗性基因（Km），得到基因元件hom::Km；通过电击转化法将hom::Km转入出发菌株替换原菌株的hom，在含卡那霉素的平板上挑取阳性转化子，通过PCR验证得到高丝氨酸脱氢酶缺陷的重组菌。发酵结果表明重组菌C.g- hom::Km -8发酵60小时赖氨酸产量达到4.7 g／L，是出发菌株谷氨酸棒杆菌ATCC 13032（0.7 g／L）的6.7倍。

Abstract: In this research the hom gene encoding for homoserine dehydrogenase was amplified from the genomic DNA of Corynebacterium glutamicum ATCC 13032. After the kanamycin-resistant gene (Km) cassette from plasmid pET28a was inserted into the center of hom, the hom::Km cassette was then electroporated into the competent cell of C. glutamicum ATCC 13032. And kanamycin-resistant clones were obtained. PCR was performed to confirm whether the Km gene was integrated into the hom gene of these clones and the recombinant strains of hom-disrupted were screened out. Fermentation results showed that the lysine yield of the hom-disrupted strain C.g-hom::Km-8 reached 4.7 g/L, which was 6.7 times that of C. glutamicum ATCC 13032.

收稿日期: 2006-07-26 出版日期: 2010-05-10

基金资助: 国家自然科学基金

通讯作者: 诸葛健 E-mail: jzhuge@sohu.com

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引用本文:

饶志明张君胜沈微方慧英诸葛健. 谷氨酸棒杆菌高丝氨酸脱氢酶编码基因hom的敲除[J]. 中国生物工程杂志, 2007, 27(1): 59-63.

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https://manu60.magtech.com.cn/biotech/CN/Y2007/V27/I1/59

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