Calpastatin (CAST) gene is closely related with meat quality in livestock and poultry. Based on the bovine and ovine mRNA sequences, the cDNA of CAST Ⅱgene in goat was amplified successfully for the first time by using RACE-PCR. Results showed that CASTⅡof goat was 2474 bp in length with an open reading frame (ORF) 1695 bp long and encoded 564 amino acids, and there were four conserved domains and one conserved seven-peptide do-main in amino acids sequences. Bioinformation analysis indicated that its secondary structures mainly were random coil and helical regions, and contained rich hydrophobic regions, certain phosphorylation sites, and protein kinase C (PKC) sites. Meanwhile, analysis of tissue expression of the gene in Tianfu meat goat demonstrated it was expressed in seven selected tissues. When the goat was of 6-month age, the highest expression was observed in longissimusdorsi, which was significantly higher than that of crureus (P<0.05) and other internal organ tissues (P<0.01). Furthermore, the expression of CAST II increased with the rise of the age and became the highest when the goat was at three-year age.
ZHAO Bo-Yang, WANG Dai-Hua, XU Gang-Yi, ZHAO Wen-Bei, ZHENG Cheng-Li
. Cloning and tissue expression of CAST transcriptⅡ in goat[J]. Hereditas(Beijing), 2011
, 33(4)
: 358
-364
.
DOI: 10.3724/SP.J.1005.2011.00358
[1] Koohmaraie M. The role of Ca2+-dependent proteases (calpains) in post mortem proteolysis and meat tenderness. Biochimie, 1992, 74(3): 239-245.
[2] Zhang LP, Ma BY, Wu JP, Fei CH, Yang L,Wan HL. Cloning and characterization of the yak gene coding for calpastatin and in silico analysis of its putative product. Acta Biochim Pol, 2010,57(1): 35-41.
[3] Barnoy S, Glaser T, Kosower NS. The role of calpastatin (the specific calpain inhibitor) in myoblast differentiation and fusion. Biochem Biophys Res Commun, 1996, 220(3): 933-938.
[4] Huang J, Forsberg NE. Role of calpain in skeletal-muscle protein degradation. Proc Nat Acad Sci USA, 1998, 95(21): 12100-12105.
[5] Killefer J, Koohmaraie M. Bovine skeletal muscle calpastatin: cloning, sequence analysis, and steady-state mRNA expression. J Anim Sci, 1994, 72(3): 606-614.
[6] Maki M, Takano E, Osawa T, Ooi T, Murachi T, Hatanaka M. Analysis of structure-function relationship of pig calpastatin by expression of mutated cDNAs in Escherichia coli. J Biol Chem, 1988, 263(21): 10254-10261.
[7] Rettenberger G, Bruch J, Fries R, Archibald AL, Hameister H. Assignment of 19 porcine type I loci by somatic cell hybrid analysis detects new regions of conserved synteny between human and pig. Mamm Genome, 1996, 7(4): 275-279.
[8] Asada K, Ishino Y, Shimada M, Shimojo T, Endo M, Kimizuka F, Kato I, Maki M, Hatanaka M, Murachi T. cDNA cloning of human calpastatin: sequence homology among human, pig, and rabbit calpastatins. J Enzym Inhib Med Chem, 1989, 3(1): 49-56.
[9] Takano J, Kawamura T, Murase M, Hitomi K, Maki M. Structure of mouse calpastatin isoforms: implications of species-common and species-specific alternative splicing. Biochem Biophys Res Commun, 1999, 260(2): 339-345.
[10] Emori Y, Kawasaki H, Imajoh S, Imahori K, Suzuki K. Endogenous inhibitor for calcium-dependent cysteine protease contains four internal repeats that could be responsible for its multiple reactive sites. Proc Natl Acad Sci USA, 1987, 84(11): 3590-3594.
[11] Lee WJ, Hatanaka M, Maki M. Multiple forms of rat calpastatin cDNA in the coding region of functionally unknown amino-terminal domain. Biochim Biophys Acta, 1992, 1129(2): 251-253.
[12] Goll DE, Thompson VF, Li HQ, Wei W, Cong JY. The calpain system. Physiol Rev, 2003, 83(3): 731-801.
[13] Croall DE, DeMartino GN. Calcium-activated neutral protease (calpain) system: structure, function, and regulation. Physiol Rev, 1991, 71(3): 813-847.
[14] Takano E, Ma H, Yang HQ, Maki M,Hatanaka M. Preference of calcium-dependent interactions between calmodulin-like domains of calpain and calpastatin subdomains. FEBS Lett, 1995, 362(1): 93-97.
[15] 许梓荣, 胡彩虹, 李卫芬. 钙蛋白酶系统的结构、活性调节及其在骨骼肌生长中的作用. 中国畜牧杂志, 2002, 38(2): 44-46.
[16] 张菊, 杜立新, 魏彩虹, 李宏滨. 绵羊CAST基因2型和4型转录本的克隆及特性分析. 遗传, 2009, 31(11): 1107-1112.
[17] 刘安芳, 朱庆, 刘益平, 张增荣. 钙蛋白酶抑制蛋白(CAST)基因在鸡不同组织和品种中的表达差异. 中国农业科学, 2007, 40(10): 2331-2335.
[18] Remignon H, Gardahaut MF, Marche G, Ricard FH. Selection for rapid growth increases the number and the size of muscle fibres without changing their typing in chickens. J Muscle Res Cell Motil, 1995, 16(2): 95-102.
[19] 曾勇庆, 王慧, 储明星. 小尾寒羊肉品理化性状及食用品质的研究. 中国畜牧杂志, 2000, 36(3): 6-8.
[20] 曾勇庆, 孙玉民, 王慧, 李同树. 青山羊肉品理化性状及其食用品质的研究. 山东农业大学学报, 1999, 30(4): 384-389.
[21] Morgan JB, Wheeler TL, Koohmaraie M, Savell JW, Crouse JD. Meat tenderness and the calpain proteolytic system in longissimus muscle of young bulls and steers. J Animl Sci, 1993, 71(6): 1471-1476.
