Ogawa T
Solar Energy Research Group, Institute of Physical and Chemical Research, Saitama, Japan.
Proc Natl Acad Sci U S A. 1991 May 15;88(10):4275-9. doi: 10.1073/pnas.88.10.4275.
A clone that transforms the RKa mutant of Synechocystis PCC6803 defective in inorganic carbon (Ci) transport to the wild-type phenotype was isolated from a cyanobacterial genomic library. The clone contained an 11.8-kilobase-pair DNA insert. Sequencing of the insert DNA in the region of the mutation in RKa revealed an open reading frame (designated as ndhB), which showed extensive amino acid sequence homology to the subunit-2 genes of NADH dehydrogenase (EC 1.6.99.3) (ndhB) of chloroplasts and mitochondria. The homology was much stronger with the chloroplast genes. Sequence analysis of the ndhB gene of RKa mutant revealed a G----A substitution that results in a Gly----Asp substitution in the deduced amino acid. A defined mutant (M55), constructed by inactivating the ndhB gene in wild-type Synechocystis, required high CO2 conditions for growth and was unable to transport CO2 and HCO3- into the intracellular Ci pool. The results indicate that the ndhB gene is required for Ci transport. Dark respiration was also depressed by the inactivation of the ndhB gene. A possible role of the ndhB gene product in the energization of Ci transport is discussed.
从蓝藻基因组文库中分离出一个克隆,该克隆可将集胞藻PCC6803中无机碳(Ci)转运缺陷的RKa突变体转化为野生型表型。该克隆包含一个11.8千碱基对的DNA插入片段。对RKa突变区域的插入DNA进行测序,发现一个开放阅读框(命名为ndhB),它与叶绿体和线粒体中NADH脱氢酶(EC 1.6.99.3)(ndhB)的亚基2基因具有广泛的氨基酸序列同源性。与叶绿体基因的同源性更强。对RKa突变体的ndhB基因进行序列分析,发现一个G→A替换,导致推导氨基酸中甘氨酸被天冬氨酸取代。通过使野生型集胞藻中的ndhB基因失活构建的一个特定突变体(M55),生长需要高CO2条件,并且无法将CO2和HCO3-转运到细胞内Ci池中。结果表明,ndhB基因是Ci转运所必需的。ndhB基因失活也会抑制暗呼吸。文中讨论了ndhB基因产物在Ci转运能量化中的可能作用。