Chen A, Hillman J D, Duncan M
Department of Oral Biology, University of Florida College of Dentistry, Gainesville 32610.
J Bacteriol. 1994 Mar;176(5):1542-5. doi: 10.1128/jb.176.5.1542-1545.1994.
The previously cloned gene for L-(+)-lactate dehydrogenase (LDH) from Streptococcus mutans was mutagenized in vitro. An Escherichia coli transformant which expressed a thermolabile LDH activity was identified. The ldh(Ts) gene was introduced into S. mutans on a suicide vector to create a heterodiploid expressing both wild-type and thermolabile LDH activities. Self-recombinants which had only one ldh gene were isolated. One of these clones expressed only the thermolabile LDH activity. This isolate grew well at 30 degrees C but did not grow at 42 degrees C under a variety of cultivation conditions, thereby proving that LDH deficiency is lethal in S. mutans in the absence of compensatory mutations.
先前克隆的变形链球菌L-(+)-乳酸脱氢酶(LDH)基因在体外进行了诱变。鉴定出一株表达热不稳定LDH活性的大肠杆菌转化体。将ldh(Ts)基因通过自杀载体导入变形链球菌,以创建一个同时表达野生型和热不稳定LDH活性的异源二倍体。分离出仅含有一个ldh基因的自重组体。其中一个克隆仅表达热不稳定LDH活性。该分离株在30℃下生长良好,但在多种培养条件下42℃时不生长,从而证明在没有补偿性突变的情况下,LDH缺陷在变形链球菌中是致死的。