首页> 中文期刊> 《畜牧兽医学报》 >野猪CAPN10基因两个剪接变异体的克隆、序列分析和表达

野猪CAPN10基因两个剪接变异体的克隆、序列分析和表达

         

摘要

In order to further study the roles of CAPN10 in muscle growth and meat quality, the study was designed to clone the cDNA sequence of Wild boar CAPN10 with RT-PCR and RACE methods, and to analyze its expression profile with semi-quantitative RT-PCR and competitive RT-PCR. Two transcript variants of CAPN10 were obtained, and the sequence analysis showed that the two polypetides were identical in the former 604 amino acids, and had the characteristic domain of calpain family, CALPAIN_CAT structure, and the catalytic triad residues in the same position. Semi-quantitative RT-PCR analysis indicated that CAPN10 mRNA of Wild boar was expressed in 12 tissues examined and the mRNA level in muscles from Yorkshire pigs at 6, 9, 12-month-old was much higher than that from crossbred pigs (P<0.05) at the same development stages. Competitive RT-PCR results indicated that the two variants were all expressed in 9 tis-sues of Wild boar examined. It is suggested that there is association between the relative amounts of CAPN10 and meat tenderness.%为了进一步研究CAPN10在肌肉生长和肉质中的作用,本研究采用RT-PCR和RACE相结合的方法克隆野猪CAPN10 cDNA并进行序列分析,采用相对定量RT-PCR和竞争性RT-PCR方法研究该基因的表达情况.获得了野猪CAPN10的2个剪接变异体,二者前604个氨基酸完全相同,在多肽链相同位置处都存在着钙蛋白酶家族的特征性结构域和催化活性中心;相对定量RT-PCR分析表明,CAPN10普遍表达于所检测的野猪12种组织内,在6、9、12月龄的肌肉组织中大白猪的表达量显著高于同一发育时期的野家杂交猪(P<0.05);竞争性RT-PCR分析表明,2个变异体都表达于所检测的野猪9种组织内.结果表明CAPN10的表达量与肌肉的嫩度可能具有一定的相关性.

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