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Title 

Transformation using in vivo and in vitro methylation in Streptomyces griseus

Authors 

Jang Yul KwakH JiangK E Kendrick

Publisher 

Wiley-Blackwell

Issue Date 

2002

Citation 

FEMS Microbiology Letters, vol. 209, no. 2, pp. 243-248

Keywords 

methylationrestriction-modification enzymestreptomyces griseustransformationbacterium transformationin vitro studyin vivo studyDNA Methylation

Abstract 

Streptomyces griseus does not readily take up foreign DNA isolated from other Streptomyces species or Escherichia coli, presumably due to its unique restriction-modification systems that function as a barrier for interspecific DNA transfer. To efficiently transform S. griseus by avoiding the restriction barriers, we methylated incoming DNA in vivo and in vitro and treated protoplasts with heat prior to transformation. Whereas heat treatment of protoplasts or methylation of the E. coli-Streptomyces shuttle vectors (pXE4 and pKK1443) did not prominently improve the transformation efficiency, HpaII methylation of the vectors from any E. coli strains tested in this study highly increased the transformation efficiency. The highest transformation efficiency was observed when the shuttle vectors were isolated from the dam, hsd strain of E. coli (GM161) and methylated by AluI and HpaII methyltransferases, and the efficiency was approximately the same as that of the vectors from S. griseus. We identified several restriction-modification systems that decrease the transformation efficiency. This research also led us to understand methylation profiles and restriction-modification systems in S. griseus.

ISSN 

0378-1097

Link 

http://dx.doi.org/10.1016/S0378-1097(02)00512-8

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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