Evaluation of the hok/sok killer locus for enhanced plasmid stability

Biotechnology and Bioengineering - Tập 44 Số 8 - Trang 912-921 - 1994
Kuo‐Wei Wu1, Thomas K. Wood1
1Department of Chemical and Biochemical Engineering, University of California at Irvine, Irvine, California 92717–2575

Tóm tắt

AbstractThe effectiveness of the hok/sok plasmid stability locus and mechanism of cloned‐gene loss was evaluated in shake‐flask cultures. Addition of the hok/sok locus dramitically increasedapparent plasmid segregational stability to the hok/sok− control. In terms of the number of generations before 10%of the population became plasmid‐free, segregational stability was increased by 11‐ to 20‐fold in different media in the absence of induction of the cloned‐gene (hok/sok+ plasmid stable for over 200 generations in all media tested). With constant expression of β‐galactosidase in the absence of an tibiotic, the segregational stability of the plasmid containing hok/sok was incresed more than 17‐ to 30‐fold when β‐galactosidase was expressed at 7–15 wt % of total cell protein. Although the hok/sok system stabilized the plasmid well infour different media (Luria‐Bertani (LB), LB glucose, M9C Trp, and a representative fedbatch medium), the ability of hok/sok to maintain the plasmid with induction of the cloned gene decreased as the complexity of the media increased. This result is better interpreted in terms of the influence of cloned‐gene expression on plasmidmaintenance; plasmid segregational stability decreased linearly as specificβ‐galactosidase activity increased. © 1994 John Wiley & Sons, Inc.

Từ khóa


Tài liệu tham khảo

10.1002/bit.260370808

Csonka L. N., 1979, Deletions generated by the transposon Tn10 in the srl recA region of the E. coli K‐12 chromosome, Genetics, 93, 321, 10.1093/genetics/93.2.321

10.1002/bit.260380917

10.1080/00986448608911387

10.1038/nbt1288-1402

10.1002/j.1460-2075.1986.tb04459.x

10.1111/j.1749-6632.1981.tb14172.x

10.1007/BF01022319

Laban A., 1981, Interplasmidic and intraplasmidic recombination in E. coli K‐12, Molec. Gen. Genet., 184, 200, 10.1007/BF00272905

10.1111/j.1462-5822.2007.00901.x

10.1021/bp00022a001

10.1007/BF02814352

10.1038/nbt1086-901

10.1002/bit.260390402

10.1111/j.1472-765X.1992.tb00693.x

10.1038/nbt0190-47

1983 Benjamin/Cummings Publishing Menlo Park CA. R. L. Rodriguez R. C. Tait Recombinant DNA techniques: an introduction

10.1002/bit.260330702

10.1007/BF00169425

10.1002/bit.260270105

10.1016/0378-1119(87)90314-3

10.1016/0022-2836(92)90714-U

10.1007/BF01027124

10.1007/BF00156608

10.1002/bit.260360902

10.1002/bit.260380410

10.1002/bit.260380811

Thông tin
Thông tin xuất bản

Biotechnology and Bioengineering

Tập 44 Số 8

912-921

Thông tin tác giả