A spontaneous runaway vector for production-scale expression of bovine somatotropin from Escherichia coli

C. Trepod; J. Mott

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A spontaneous runaway vector for production-scale expression of bovine somatotropin from Escherichia coli

机译：自发失控载体，用于大肠杆菌中牛生长激素的生产规模表达

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An Escherichia coli expression vector was constructed for the production-scale fermentation of recombinant bovine somatotropin (rBST). Gene expression is regulated by a spontaneous increase in copy number at a constant low temperature without the need for an external inducer. This vector, designated pURA-4, contains the ampicillin resistance gene, the replication origin from pBR322, the R1 temperature-inducible runaway replicon, and a gene encoding rBST. Optimized rBST expression levels of >35% total cell protein were achieved at a constant 28°C. Shake-flask analysis of pURA-4 shows that the copy number spontaneously increases approximately 6-fold during rBST production. Investigation into the mechanism of pURA-4 spontaneous runaway shows that the increase in copy number is directed by the pBR322 ori and not by the R1 replicon. Although the R1 temperature-inducible replicon does not mediate spontaneous runaway, it does have a positive effect on rBST expression. Copy number analysis also confirmed the stability of pURA-4 spontaneous runaway from the shake-flask scale through the production scale.

机译：构建了大肠杆菌表达载体，用于重组牛生长激素（rBST）的生产规模发酵。基因表达受到恒定恒定低温下拷贝数的自发增加的调节，而无需外部诱导剂。命名为pURA-4的该载体包含氨苄青霉素抗性基因，来自pBR322的复制起点，R1温度可诱导的失控复制子和编码rBST的基因。在恒定的28°C下，rBST的最佳表达水平>总细胞蛋白> 35％。 pURA-4的摇瓶分析表明，在rBST生产过程中，拷贝数自发增加了约6倍。对pURA-4自发失控机理的研究表明，拷贝数的增加是由pBR322 ori而不是R1复制子引起的。尽管R1温度诱导型复制子不介导自发失控，但它确实对rBST表达具有积极作用。拷贝数分析还证实了从摇瓶规模到生产规模，pURA-4自发失控的稳定性。

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《Applied Microbiology and Biotechnology》 |2002年第1期|84-88|共5页
作者
C. Trepod; J. Mott;
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Pharmacia Infectious Diseases Biology 7263–209–713 Kalamazoo MI 49007 USA;

Pharmacia Infectious Diseases Biology 7263–209–713 Kalamazoo MI 49007 USA;

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1. A spontaneous runaway vector for production-scale expression of bovine somatotropin from Escherichia coli [J] . Trepod CM., Mott JE. Applied Microbiology and Biotechnology . 2002,第1期

机译：自发失控载体，用于大肠杆菌中牛生长激素的生产规模表达
2. High-level expression of bovine rotavirus VP4 protein in Escherichia coli using a hepatitis B core antigen vector [J] . Nazlina Ibrahim, Malcolm A.McCrae Biotechnology Letters . 2001,第8期

机译：乙型肝炎核心抗原载体在牛大肠杆菌中轮状表达VP4蛋白
3. A spontaneous direct repeat deletion in the pGEX fusion vector decreases the expression level of recombinant proteins in Escherichia coli [J] . Borgi I, Gargouri A Protein Expression and Purification . 2008,第1期

机译：pGEX融合载体中的自发直接重复缺失会降低重组蛋白在大肠杆菌中的表达水平
4. Heterologous expression of bovine lactoferricin in Escherichia coli [C] . The 2nd International Conference on Bioinformatics and Biomedical Engineering(iCBBE 2008)(第二届生物信息与生物医学工程国际会议）论文集 . 2008

机译：牛乳铁蛋白在大肠杆菌中的异源表达
5. Development of low-copy expression vectors derived from the F plasmid of Escherichia coli. [D] . Jones, Kristala Lanett. 1999

机译：衍生自大肠杆菌F质粒的低拷贝表达载体的开发。
6. Effect of nucleotide sequences directly downstream from the AUG on the expression of bovine somatotropin in E. coli. [O] . C S Tomich, E R Olson, M K Olsen, 1989

机译：直接在AUG下游的核苷酸序列对牛生长激素在大肠杆菌中表达的影响。
7. Effect of nucleotide sequences directly downstream from the AUG on the expression of bovine somatotropin in E. coli. [O] . Tomich, C S, Olson, E R, Olsen, M K, 1989

机译：直接在AUG下游的核苷酸序列对牛生长激素在大肠杆菌中表达的影响。

A spontaneous runaway vector for production-scale expression of bovine somatotropin from Escherichia coli

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