Cloning and expression of two new prolactin-related proteins, prolactin-related protein-VIII and -IX, in bovine placenta
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Cloning and expression of two new prolactin-related proteins, prolactin-related protein-VIII and -IX, in bovine placenta Koichi Ushizawa1, Toru Takahashi1, Misa Hosoe1, Kanako Kaneyama1,3 and Kazuyoshi Hashizume*2 Address: 1Reproductive Biology and Technology Laboratory, Developmental Biology Department, National Institute of Agrobiological Sciences, 2 Ikenodai, Tsukuba, Ibaraki 305-8602, Japan, 2Department of Veterinary Medicine, Faculty of Agriculture, Iwate University, 3-18-8 Ueda, Morioka, Iwate 020-8550, Japan and 3Department of Technology, National Livestock Breeding Center, 1 Odakurahara, Odakura, Nishigo, Fukushima 9618511, Japan Email: Koichi Ushizawa - [email protected]; Toru Takahashi - [email protected]; Misa Hosoe - [email protected]; Kanako Kaneyama - [email protected]; Kazuyoshi Hashizume* - [email protected] * Corresponding author
Published: 07 December 2005 Reproductive Biology and Endocrinology 2005, 3:68
doi:10.1186/1477-7827-3-68
Received: 07 October 2005 Accepted: 07 December 2005
This article is available from: http://www.rbej.com/content/3/1/68 © 2005 Ushizawa et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: Prolactin-related proteins (PRPs) are specific proteins of the growth hormone/ prolactin (GH/PRL) family in bovine placenta. This study reports the identification and sequencing of a full-length cDNA for two new members of bovine PRPs, bPRP-VIII and -IX, and their localization and quantitative expression in bovine placenta. Methods: New bPRP-VIII and -IX were identified from bovine placentome. Localization and quantitative gene expression in the placenta were respectively investigated by in situ hybridization and real-time RT-PCR methods. Recombinant proteins of these genes were produced by a mammalian HEK293 cell expression system. Results: Full-length bPRP-VIII and -IX cDNA were respectively cloned with 909 and 910 nucleotide open-reading-frames corresponding to proteins of 236 and 238 amino acids. The predicted bPRP-VIII amino acid sequence shared about 40 to 70% homology with other bPRPs, and bPRP-IX had about 50 to 80 % homology of others. The two new bPRPs were detected only in the placenta by RT-PCR. mRNA was primarily expressed in the cotyledon and intercotyledonary tissues throughout gestation. An in situ hybridization analysis revealed the presence of bPRP-VIII and -IX mRNA in the trophoblastic binucleate and/or trinucleate cells. bPRP-VIII mRNA was observed in the extra-embryonic membrane on Day 27 of gestation, however, no bPRP-IX mRNA was observed in the extra-embryonic membrane in the same stage of pregnancy by quantitative real-time RT-PCR analysis. Both new bPRP genes were possible to translate a mature protein in a mammalian cell expression sys
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