三疣梭子蟹Sox基因HMG盒的克隆分析

利用能扩增人类SRY基因HMG盒的一对简并引物,分别对三疣梭子蟹雌雄个体基因组DNA进行扩增,均得到216 bp和约400 bp的两条带,不存在性别差异,通过PCR-SSCP分析,未发现雌蟹或雄蟹所特有的Sox基因。对216 bp的带进行克隆测序得到6个Sox基因,分别命名为PTSox21、PTSox12、PTSox11a、PTSox11b、PTSox11c和PTSox4。其中,PTSox21、PTSox12和PTSox4氨基酸序列与人类Sox21、Sox12和Sox4基因的同源性分别为90%、66%和63%,PTSox11a、PTSox11b和PTSox11c氨基酸序列均与人类Sox11基因有63%的同源性。此外,PTSox11a和PTSox11b的氨基酸序列之间的同源性达到了98.6%,只在第44位氨基酸残基不同。与其他物种Sox基因氨基酸序列的同源性比较发现,PTSox21与饰纹姬蛙Sox2a有92%的同源性,而PTSox12、PTSox11a、PTSox11b和PTSox11c均与意大利蜜蜂的Sox1有73%的同源性,PTSox4与意大利蜜蜂的Sox1有72%的同源性。

Cloning and Sequencing of Sox Gene HMG-box in Crab Portunus trituberculatus

Here the cloning sequence of Sox gene HMG-box in crab Portunus trituberculatus was obtained by PCR using degenerate nucleotide primers that was designed based on the amino acid sequence of human SRY gene.Two amplification bands with the length of 216 bp and 400 bp were observed and there was no difference between male and female crabs.Meanwhile the fragment of 216 bp was analysed by PCR-SSCP,and the male-or female-specific Sox gene was not found.Six Sox gene HMG-boxes were cloned from the 216 bp fragment,which were defined as PTSox21,PTSox12,PTSox11a,PTSox11b,PTSox11c and PTSox4.The identity of amino acid sequences encoded by PTSox21,PTSox12 and PTSox4 to human Sox21,Sox12 and Sox4 HMG-box was 90%,66% and 63%,respectively;the encoded amino acid sequences by PTSox11a,PTSox11b and PTSox4 exhibited 63%,63%,and 63% of sequence homology with human Sox11 HMG-box,respectively.In addition,the homology of amino acid sequences encoded by PTSox11a and PTSox11b to each other was 98.6%,and difference only in the 44th amino acid residue.And the highest identity of amino acid sequence to the six Sox genes from other animals was PTSox21 which exhibited 92% of sequence homology with Microhyla ornata Sox2a;PTSox12,PTSox11a,PTSox11b and PTSox11c exhibited 73% with Apis mellifera Sox1;PTSox4 exhibited 72% with Apis mellifera Sox1.