Gill S S, Ross L S
Department of Entomology, University of California, Riverside 92521.
Arch Biochem Biophys. 1991 Nov 15;291(1):92-9. doi: 10.1016/0003-9861(91)90109-v.
Using the polymerase chain reaction (PCR) a 0.8-kb product was amplified from cDNA made from the midgut and Malpighian tubules of fifth instar larvae of Helicoverpa virescens. This 0.8-kb PCR product was then used to isolate a clone of the B subunit of the V-type ATPase from a cDNA library made from the same tissues. The cDNA clone encodes for a protein of 55 kDa which shows very high amino acid homology to other known B subunits of V-type ATPases. The transcript size of the B subunit in the midgut of H. virescens was 2.3 kb, and a transcript of identical size was also detected in the Malpighian tubules. Northern blot analysis revealed the presence of a homologous transcript of 2.6 kb in the midgut of Manduca sexta and PCR analysis also confirmed the presence of such a transcript in the Malpighian tubules and the nervous system of M. sexta, and in the midgut Malpighian tubules of Culex quinquefasciatus. The presence of the V-type ATPase in the Malpighian tubules of lepidopteran insects suggests that the transport of ions across the cell membrane in this tissue is also probably driven by a similar process as that observed in the midgut of these insects.
利用聚合酶链反应（PCR）从绿铃夜蛾五龄幼虫中肠和马氏管制备的cDNA中扩增出一个0.8 kb的产物。然后，该0.8 kb的PCR产物被用于从同一组织制备的cDNA文库中分离V型ATP酶B亚基的一个克隆。该cDNA克隆编码一个55 kDa的蛋白质，它与其他已知的V型ATP酶B亚基具有非常高的氨基酸同源性。绿铃夜蛾中肠中B亚基的转录本大小为2.3 kb，在马氏管中也检测到相同大小的转录本。Northern印迹分析显示，烟草天蛾中肠存在一个2.6 kb的同源转录本，PCR分析也证实烟草天蛾的马氏管和神经系统以及致倦库蚊的中肠和马氏管中存在这样一个转录本。鳞翅目昆虫马氏管中存在V型ATP酶表明，该组织中离子跨细胞膜的转运可能也由与这些昆虫中肠中观察到的类似过程驱动。
