Molecular cloning of P-type ATPases on intracellular membranes of the marine alga Heterosigma akashiwo |
| |
Authors: | Wada Masato Shono Mariko Urayama Osamu Satoh Shinobu Hara Yukichi Ikawa Yoji Fujii Tadashi |
| |
Institution: | (1) Institute of Biological Sciences, University of Tsukuba, 305, Ibaraki, Tsukuba City, Japan;(2) Department of Biochemistry, Tokyo Medical & Dental University, School of Medicine, 1-5-45 Yushima Bunkyo-ku, 113 Tokyo, Japan |
| |
Abstract: | Two cDNA clones (HAA13 and HAA1) which include conserved regions of genes of P-type ATPases were isolated from the marine alga Heterosigma akashiwo by a method that included the polymerase chain reaction. The longer cDNA (3286 bp), HAA13, consisted of an open reading frame that encoded a 106 kDa polypeptide of 977 amino acids with several possible transmembrane domains and conserved regions of eukaryotic P-type ATPases. One transmembrane domain had a leucine zipper structure. HAA1 was not a full-length gene (2054 bp) and lacked the 5 region, but it also included the conserved regions and putative transmembrane domains. Antibodies against the polypeptides encoded by HAA13 and HAA1 that have been expressed in Escherichia coli reacted with 100 kDa and 95 kDa polypeptides, respectively, on intracellular membranes of H. akashiwo cells. Immunostaining of H. akashiwo cells revealed that the HAA13 antigen was distributed on membranes around chloroplasts and the HAA1 antigen was located on small vesicles. |
| |
Keywords: | P-type ATPase PCR marine alga Heterosigma akashiwo |
本文献已被 SpringerLink 等数据库收录! |
|