Isolation and cDNA Cloning of Ovarian Cortical Rod Protein in Kuruma Prawn Marsupenaeus japonicus(Crustacea: Decapoda: Penaeidae)(Reproduction Biology)

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概要

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• Two cortical rod proteins having molecular weights of 28.6 kDa and 30.5 kDa were isolated from mature ovary of Marsupenaeus japonicus using gel filtration and reversed-phase HPLC. Analysis of the N-terminal amino acid sequence of the 28.6 kDa molecule revealed that amino acid residued 1-21 corresponded to residues 9-29 of the 30.5 kDa molecule. Examination of homology using BLAST showed that 21 amino acids out of 29 residues of the 28.6 kDa molecule, and 14 out of 29 residues of the 30.5 kDa molecule were identical to that of the ovarian cortical rod proteins of Penaeus semisulcatus. Positive immunohistochemical reaction to antiserum raised against the 28.6 kDa protein was observed on cortical rods forming around the periphery of oocytes at the maturation stages. Western blotting analyses revealed that both the 28.6 kDa and 30.5 kDa molecules stained with the anti-28.6 kDa antiserum. Furthermore, the 28.6 kDa and 30.5 kDa proteins were both glycosylated, as evidensed by positive carbohydrate staining using Concanavalin A and production of positive PAS reaction. These results indicate that the cortical rods are comprised of the 28.6 kDa and 30.5 kDa molecules. We subsequently cloned two full-length cDNAs based on the N-terminal sequences of the 28.6 kDa and 30.5 kDa molecules. The open reading frame of 28.6 kDa and 30.5 kDa encoded 276 amino acid residues. Comparison analysis of the two cDNAs revealed that the location of the processing site and sequence of signal peptides differed, indicating that the two cDNAs are products of two separate genes and encode the 28.6 kDa molecule and 30.5 kDa molecule, respectively. Both proteins possessed one potential N-linked glycosylation site. It is considered that both molecules are components of the cortical rods, forming a jelly layer after fertilization.

• 2004-11-25

著者

• JASMANI Safiah

  Japan International Research Center for Agricultural Sciences

• WILDER Marcy

  Japan International Research Center for Agricultural Sciences

• KIM YI

  Graduate School of Agricultural and Life Sciences, The University of Tokyo

• KIM Yi

  Department of Aquatic Life Medicine, College of Fisheries Science, Pukyoung National University

• Aida Katsumi

  Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University

• Tsutsui Naoaki

  Japan International Research Center for Agricultural Sciences

• Aida K

  Department Of Aquatic Bioscience Graduate School Of Agricultural And Life Sciences University Of Tok

• Wilder M

  Japan International Research Center For Agricultural Sciences

• Wilder Marcy

  Department Of Fisheries Faculty Of Agriculture The University Of Tokyo

• Wilder Marcy

  Japan International Research Center For Agricultural Sciences Ministry Of Agriculture Forestry And F

• Kawazoe Ichiro

  Department of Aquatic Bioscience, Gradualte School of Agricultural and Life Sciences, The University

• Kawazoe Ichiro

  Japan International Research Center for Agricultural Sciences

• Kawazoe Ichiro

  Japan International Research Center For Agriculture Sciences

• Kawazoe I

  Univ. Tokyo Tokyo Jpn

• Kawazoe Ichiro

  Institute For Research And Development Taiyo Fishery Co. Ltd.

• Aida Katsumi

  Department Of Aquatic Bioscience Graduate School Of Agricultural And Life Sciences The University Of

• Aida Katsumi

  Department Of Aquatic Bioscience Graduate School Of Agricultural And Sciences The University Of Toky

• Katayama Akira

  Department Of Biochemistry And Molecular Biology Nippon Medical School

• Tsutsui Naoaki

  Department Of Applied Biological Chemistry Graduate School Of Agricultural And Life Sciences The Uni

• Tsutsui Naoaki

  Department Of Molecular Biology Ocean Research Institute The University Of Tokyo:department Of Appli

• Jasmani S

  Japan International Research Center For Agricultural Sciences

• Kim Yi

  Department Of Aquatic Bioscience Graduate School Of Agricultural And Life Sciences University Of Tok

• Wilder Marcy

  Japan International Res. Center For Agricultural Sci. Ibaraki Jpn

• Wilder Marcy

  Japan Int'l Research Center For Agricultural Sciences

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