Centre for Biotechnology in Agriculture Research, Division of Genetics & Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
Fish Shellfish Immunol. 2012 Jan;32(1):161-9. doi: 10.1016/j.fsi.2011.11.006. Epub 2011 Nov 18.
Caspase 3c (MrCasp3c) was sequenced from the freshwater giant prawn Macrobrachium rosenbergii using Illumina Solexa Genome Analyzer Technique. MrCasp3c consisted of 2080 bp nucleotide encoded 521 polypeptide with an estimated molecular mass of 59 kDa. MrCasp3c sequence contains caspase family p20 domain profile and caspase family p10 domain profile at 236-367 and 378-468 respectively. The quantitative real time PCR analysis revealed a broad expression of MrCasp3c with the highest expression in haemocyte and the lowest in stomach. The expression of MrCasp3c after challenge with the infectious hypodermal and haematopoietic necrosis virus (IHHNV) was tested in haemocyte. In addition, MrCasp3c was expressed in Escherichia coli by prokaryotic expression plasmid pMAL-c2x. The enzyme activity of MrCasp3c was also found to be up-regulated by IHHNV in haemocyte and hepatopancreas tissues. This study suggested that MrCasp3c may be an effector caspase associated with the induction of apoptosis which is potentially involved in the immune defence of M. rosenbergii.
使用 Illumina Solexa 基因组分析仪技术，从淡水巨型对虾（Macrobrachium rosenbergii）中测序得到 Caspase 3c（MrCasp3c）。MrCasp3c 由 2080bp 核苷酸编码 521 个多肽组成，估计分子量为 59kDa。MrCasp3c 序列包含半胱氨酸蛋白酶家族 p20 结构域和半胱氨酸蛋白酶家族 p10 结构域，分别位于 236-367 和 378-468 位。定量实时 PCR 分析显示 MrCasp3c 广泛表达，在血细胞中表达最高，在胃中表达最低。在血细胞中检测到感染性皮下和造血坏死病毒（IHHNV）后 MrCasp3c 的表达。此外，MrCasp3c 通过原核表达质粒 pMAL-c2x 在大肠杆菌中表达。还发现 IHHNV 在血细胞和肝胰腺组织中上调 MrCasp3c 的酶活性。本研究表明，MrCasp3c 可能是一种与细胞凋亡诱导相关的效应半胱氨酸蛋白酶，可能参与了 M.rosenbergii 的免疫防御。
