%0 Journal Article %T Highly efficient transformation with plasmid DNA in Klebsiella pneumoniae
外源载体高效转化肺炎克雷伯菌的新途径 %A ZHENG Yan %A LIU Xi-peng %A LIU Jian-hua %A
郑艳 %A 刘喜朋 %A 刘建华 %J 微生物学报 %D 2007 %I %X Attempts to transform Klebsiella pneumoniae resulted in very low efficiencies because of capsule polysaccharide (CPS). It was reported that some chelating agents could reduce CPS production and improve transformation efficiency. These methods mentioned above could not improve transformation efficiency apparently by incorporating such agents to liquid medium. However, this method introduces a simple way for efficient transformation of K. pneumoniae. In this method, K. pneumoniae strains NTUH-K2044 and magA(-) mutant are envolved as recipients. The plasmids used in this way are composed of pIP843T, pIP843TdhaB, pIP843TdhaT with different sizes. The sole critical step is to harvest bacteria on LB plates to prepare competent cells. 150 +/- 10, 1.3 x 10(3) +/- 100, 2 x 10(5) +/- 300, and 3.4 x 10(7) +/- 500 transformants were obtained per microgram plasmid DNA with NTUH-K2044 liquid cells, magA(-) liquid cells, NTUH-K2044 solid cells, and magA(-) solid cells, respectively. The number of transformants per microg DNA obtained by electroplating solid cells is at least 10(3) fold higher than that of transformants with liquid-cultured bacteria. This method will benefit gene manipulation and genetic study in K. pneumoniae. %K Klebsiella pneumoniae
电转化 %K 转化效率 %U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=C0C8980C7B2A2DD3&yid=A732AF04DDA03BB3&vid=F4B561950EE1D31A&iid=E158A972A605785F&sid=569BDAA4FEA0F7F9&eid=94655B9881133A28&journal_id=0001-6209&journal_name=微生物学报&referenced_num=0&reference_num=12