DNA Taken into Bacillus subtilis Competent Cells by Lysed-Protoplast Transformation Is Not ssDNA but dsDNA(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)
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概要
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Competent Bacillus subtilis incorporates whole-genome DNA (4215kb) from the protoplast lysate of B. subtilis subtilis [Akamatsu, T. and Taguchi, H., Biosci. Biotechnol. Biochem., 65, 823-829 (2001)]. A continuous incorporated DNA is longer than 1500kb [J. Biosci. Bioeng., 101, 257-262 (2006)]. Whether the incorporated DNA is single-stranded (ssDNA) or double-stranded DNA (dsDNA) has been studied by examining the transforming activity of the incorporated DNA. B. subtilis BEST7027 was used as the donor strain, which has a heterologous region consisting of the 145kb region of the Synechocystis sp. PCC6803 genome and erm gene. The donor DNA was transferred to a wild-type or a recA recipient strain (AYG2 or SYN9), and protoplast lysate was prepared from the transformants and used as the donor DNA source for the second recipient strain (AU1 or AV1). The intergenote region showed a significant transforming activity. When DNase I was added to both cells collected from the first transformation mixture and the following protoplastization, the result was similar to that obtained without DNase I. All of the observations strongly suggest that the incorporated DNA is dsDNA, and the transformation of competent B. subtilis by DNA in protoplast lysate is different from that by purified DNA taken up conventionally.
- 公益社団法人日本生物工学会の論文
- 2006-04-25
著者
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Taguchi Hisataka
Department of Applied Microbial Technology, Faculty of Biotechnology and Life Science, Sojo Universi
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Saito Yukiko
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Akamatsu Takashi
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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Taguchi Hisataka
Department Of Applied Microbial Technology Faculty Of Biotechnology And Life Science Sojo University
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