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Molecular Cloning, Bioinformatics Analysis and Expression Profiling of a Gene Encoding Vacuolar-type $H^+-ATP$ Synthetase (V-ATPase) c Subunit from Bombyx mori  

Lu, Peng (Institute of Life Sciences, Jiangsu University)
Chen, Keping (Institute of Life Sciences, Jiangsu University)
Yao, Qin (Institute of Life Sciences, Jiangsu University)
Yang, Hua-Jun (Institute of Life Sciences, Jiangsu University)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.15, no.2, 2007 , pp. 115-122 More about this Journal
Abstract
As the genome of B.mori is available in GenBank and the EST database of B.mori is expanding, identification of novel genes of B.mori is conceivable by data-mining techniques. We used the in silico cloning method to get the vacuolar-type $H^+-ATP$ synthetase (V-ATPase) c subunit (16 kDa proteolipid subunit) gene of B.mori and analysed with bioinformatics tools. The result was confirmed by RT-PCR and sequencing. The V-ATPase c subunit cDNA contains a 468 bp ORF. The ORF encoded a 155-residue protein that showed extensive homology with V-ATPase c subunits from other 15 species and contained four membrane-spanning helices. Tissue expression pattern analysis revealed that V-ATPase c expressed strongly in Malpighian tubules, not in fat body. This gene has been registered in GenBank under the accession number EU082222.
Keywords
16 kDa proteolipid; RT-PCR; sequencing; tissue expression;
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1 Dunlop, J., Jones, P. C. and Finbow, M. E. (1995) Membrane insertion and assembly of ductin: a polytopic channel with dual orientations. Embo. J. 14, 3609-16
2 Galli, T., McPherson, P. S. and De Camilli, P. (1996) The V0 sector of the V-ATPase, synaptobrevin, and synaptophysin are associated on synaptic vesicles in a Triton X-100-resistant, freeze-thawing sensitive, complex. J. Biol. Chem. 271, 2193-8   DOI
3 Pali, T., Finbow, M. E. and Marsh, D. (2006) A divalent-ion binding site on the 16-kDa proton channel from Nephrops norvegicus-revealed by EPR spectroscopy. Biochimica. et Biophysica. Acta. 1758, 206-212   DOI   ScienceOn
4 Saitou, N. and Nei, M. (1987) The neighbor-joining method:a new method for reconstructing phylogenetic trees. Mol. BIol. Evol. 4, 406
5 Stevens, T. H. and Forgac, M. (1997) Structure,function and regulation of the vacuolar (H+)-ATPase. Annual Review of Cell and Developmental Biology 13, 779-808   DOI   ScienceOn
6 Harvey, W. J. and Nelson, N. (1992) (eds)V-ATPases. J. Exp. Biol. 172, 1-485
7 Nelson, N. J. (1989) Structure,molecular,genetics and evolution of vacuolar H+-ATPase. J. Bioenerg. Biomembr. 21, 553
8 Wilkens, S., Inoue, T. and Forgac, M. (2004) Three-dimensional structure of the vacuolar ATPase. Localization of subunit H by difference imaging and chemical cross-linking. J. Biol. Chem. 279, 41942-9   DOI   ScienceOn
9 Zhang, J., Feng, Y. and Forgac, M. (1994) Proton conduction and bafilomycin binding by the V0 domain of the coated vesicle V-ATPase. J. Biol. Chem. 269, 23518-23
10 Zhang, J., Myers, M. and Forgac, M. (1992) Characterization of the V0 domain of the coated vesicle (H+)-ATPase. J. Biol. Chem. 267, 9773-9778
11 Finbow, M. E., John, S., Kam, E., Apps, D. K. and Pitts, J. D. (1993) Disposition and orientation of ductin (DCCD-reactive vacuolar H(+)-ATPase subunit) in mammalian membrane complexes. Exp. Cell Res. 207, 261-70   DOI   ScienceOn
12 Finbow, M. E. and Harrison, M. A. (1997) The vacuolar H+- ATPase: a universal proton pump of eukaryotes. Biochem. J. 324(Pt 3), 697-712   DOI
13 Bairoch, A., Bucher, P. and Hofmann, K. (1997) The PROSITE database. Nucleic Acids Research 25, 217-221   DOI   ScienceOn
14 Nishi, T. and Forgac, M. (2002) The vacuolar (H+)-ATPases-- nature's most versatile proton pumps. Nat. Rev. Mol. Cell Biol. 3, 94-103   DOI   ScienceOn
15 Rost, B. (1996) PHD:predicting one-dimensional protein structure by profile based neurl networks. Methods in Enzymology 266, 525-539   DOI
16 Nelson, N. and Klionsky, D. J. (1996) Vacuolar H(+)-ATPase: from mammals to yeast and back. Experientia 52, 1101-10   DOI   ScienceOn
17 Sun, S. Z., Xie, X. S. and Stone, D. K. (1987) Isolation and reconstitution of the dicyclohexylcarbodiimide-sensitive proton pore of the clathrin-coated vesicle proton translocating complex. J. Biol. Chem. 262, 14790-14794
18 Finbow, M. E. and Meagher, L. (1992) Connexins and the vacuolar proteolipid-like 16-kDa protein are not directly associated with each other but may be components of similar or the same gap junctional complexes. Exp. Cell Res. 203, 208-4
19 Fillingame, R. H. (1996) Membrane sectors of F- and V-type H+-transporting ATPases. Curr. Opin. Struct. Biol. 6, 491- 498   DOI   ScienceOn
20 Finbow, M. E., Harrison, M. and Jones, P. (1995) Ductin--a proton pump component, a gap junction channel and a neurotransmitter release channel. Bioessays 17, 247-55   DOI   ScienceOn
21 Forgac, M. (1999) Structure and properties of the clathrincoated vesicle and yeast vacuolar V-ATPases. J. Bioenerg. Biomembr. 31, 57-65   DOI   ScienceOn
22 Kumar, S. K., Tamura, K., Jakobsen, I. B. and Nei, M. (2001) MEGA2:molecular evolutionary genetics analysis software. Bioinformatics 17, 1244-1245   DOI   ScienceOn
23 Takase, K., Kakinuma, S., Yamato, I., Konishi, K., Igarashi, K. and Kakinuma, Y. (1994) Sequencing and characterization of the ntp gene cluster for vacuolar-type Na(+)-translocating ATPase of Enterococcus hirae. J. Biol. Chem. 269, 11037- 44
24 Birman, S., Meunier, F., Lesbats, B., LeCaer, J., Rossier, J. and Lsrael, M. (1990) A 15 kDa proteolipid found in mediatophore preparations from Torpedo electric organ presents high sequence homology with the bovine chromaffin granule protonophore. FEBS Lett. 261, 303-306   DOI   ScienceOn
25 Higgins, D., Thompson, J., Gibson, T., Thompson, J. D., Higgins, D. G. and Gibson, T. J. (1994) CLUSTAL W:improving the sensitivity of progressive multiple sequence alignment through sequence weighting,position-specific gap penalties and weight matrix choice. Nucleic Acids Research 22, 4673- 4680   DOI
26 Schwede, T., Kopp, J., Guex, N. and Peitsch, M. C. (2003) SWISS-MODEL:an automated protein homology-modeling server. Nucleic Acids Research 31, 3381-3385   DOI   ScienceOn
27 Florea, L., Hartzell, G., Zhang, Z., Rubin, G. M. and Miller, W. (1998) A computer program for aligning a cDNA sequence with a genomic DNA sequence. Genome Res. 8, 967-974   DOI
28 Guex, N. and Peitsch, M. C. (1997) SWISS-MODEL and the Swiss-Pdbviewer:An environment for compatative protin modelling. Electrophoresis 18, 2714-2723   DOI   ScienceOn