Gill S S, Chu P B, Smethurst P, Pietrantonio P V, Ross L S
Department of Entomology, University of California, Riverside 92521, USA.
Arch Insect Biochem Physiol. 1998;37(1):80-90. doi: 10.1002/(SICI)1520-6327(1998)37:1<80::AID-ARCH10>3.0.CO;2-6.
Using conserved amino acid sequences for the design of oligonucleotide primers, we isolated cDNA clones for two subunits of the V-ATPase from the midgut and Malpighian tubules of Aedes aegypti larvae. The 3.1 kb cDNA of the A subunit of the peripheral catalytic V1 sector codes for a protein of 68.6 kDa. The protein contains conserved motifs, including an ATP/GTP binding site, found in all other A subunits. Southern analysis using the A subunit as a probe suggests the presence of only a single copy of gene in the Aedes aegypti. The 0.85 kb cDNA of the c subunit of the membrane H+ conducting V0 sector codes for a protein of kDa. This protein has four transmembrane domains and contains a conserved glutamic acid that serves as the binding site for dicyclohexylcarbodiimide. Southern analysis using the c subunit as a probe suggests the presence of more than one related gene in the genome of Aedes aegypti. Pileup analysis of various A and c subunits shows that these subunits fall into distinct clusters, including one in which all arthropod proteins are clustered.
利用保守氨基酸序列设计寡核苷酸引物，我们从埃及伊蚊幼虫的中肠和马氏管中分离出V - ATP酶两个亚基的cDNA克隆。外周催化V1扇区A亚基的3.1 kb cDNA编码一个68.6 kDa的蛋白质。该蛋白质含有保守基序，包括在所有其他A亚基中都存在的ATP / GTP结合位点。用A亚基作为探针进行Southern分析表明，埃及伊蚊中该基因只有一个拷贝。膜H⁺传导V0扇区c亚基的0.85 kb cDNA编码一个kDa的蛋白质。该蛋白质有四个跨膜结构域，并含有一个保守的谷氨酸，作为二环己基碳二亚胺的结合位点。用c亚基作为探针进行Southern分析表明，埃及伊蚊基因组中存在一个以上相关基因。对各种A和c亚基的比对分析表明，这些亚基分为不同的簇，包括一个所有节肢动物蛋白质都聚集在一起的簇。
