Shoemaker N B, Wang G R, Salyers A A
Department of Microbiology, University of Illinois, Urbana 61801, USA.
J Bacteriol. 1996 Jun;178(12):3594-600. doi: 10.1128/jb.178.12.3594-3600.1996.
NBU1 is a 10.3-kbp integrated Bacteroides element that can be induced to excise from the chromosome and can be mobilized to a recipient by trans-acting functions provided by certain Bacteroides conjugative transposons. The NBU1 transfer intermediate is a covalently closed circle, which is presumed to be the form that integrates into the recipient genome. We report here that a 2.4-kbp segment of NBU1 was all that was required for site-specific integration into the chromosome of Bacteroides thetaiotaomicron 5482. This 2.4-kbp region included the joined ends of the NBU1 circular form (attN1) and a single open reading frame, intN1, which encoded the integrase. Previously, we had found that NBU1 integrates preferentially into a single site in B. thetaiotaomicron 5482. We have now shown that the NBU1 target site is located at the 3' end of a Leu-tRNA gene. The NBU1 integrase gene, intN1, was sequenced. The predicted protein had little overall amino acid sequence similarity to any proteins in the databases but had limited carboxy-terminal similarity to the integrases of lambdoid phages and to the integrases of the gram-positive conjugative transposons Tn916 and Tn1545. We also report that the intN1 gene is expressed constitutively.
NBU1是一个10.3千碱基对的整合型拟杆菌元件,它可被诱导从染色体上切除,并可通过某些拟杆菌接合转座子提供的反式作用功能转移至受体细胞。NBU1转移中间体是一个共价闭合环,推测其为整合到受体基因组中的形式。我们在此报告,NBU1的一个2.4千碱基对片段是将其位点特异性整合到嗜热栖热放线菌5482染色体中所必需的全部元件。这个2.4千碱基对区域包括NBU1环状形式的连接末端(attN1)和一个单一的开放阅读框intN1,该阅读框编码整合酶。此前,我们发现NBU1优先整合到嗜热栖热放线菌5482中的一个单一位点。我们现在已经表明,NBU1靶位点位于亮氨酸转运RNA基因的3'端。对NBU1整合酶基因intN1进行了测序。预测的蛋白质与数据库中的任何蛋白质在整体氨基酸序列上几乎没有相似性,但在羧基末端与λ样噬菌体的整合酶以及革兰氏阳性接合转座子Tn916和Tn1545的整合酶有有限的相似性。我们还报告说,intN1基因是组成型表达的。