应用抑制性差减杂交技术 ( SSH)克隆两种不同小鼠胸腺基质细胞的差异表达基因 ,获得新基因片段 C55.通过 Gen Bank检索及 RT- PCR扩增出一个全长 1 .4kb的 c DNA.杂交分析认为它是一个完整的 c DNA序列 .c DNA序列分析表明 ,它拥有一个 636bp的开放读码框架 ,编码 2 1 2个氨基酸 .同源序列比较发现 ,它编码一个肌动蛋白相关蛋白的新成员 ,该序列与多种已知的肌动蛋白相关蛋白 SM2 2 α及其同源蛋白在氨基酸水平上有 62 %～ 95%的同源性 .Northern杂交分析显示 ,该基因 m RNA转录本在两种不同胸腺基质细胞中的表达存在显著差异 . RT- PCR分析显示 ,该基因特异表达于小鼠淋巴相关组织中 ,而在非淋巴组织中无表达 .
Abstract
A 200 bp novel gene fragment C55 was obtained from a mouse thymic stromal cell subtracted cDNA library by using suppression subtractive hybridization (SSH).GenBank database search and RT PCR amplification revealed a full length cDNA of 1 388 bp,which was confirmed by Northern blotting.The full length cDNA contains an open reading frame of 636 bp,encoding a predicted protein of 212 amino acids,which showed 62%～95% sequence identity with other arthropod and vertebrate SM22α.Northern blotting demonstrated differential expression of the mRNA transcript in the two thymic stromal cell lines.RT PCR analysis showed that the newly cloned gene is specifically expressed in lymphoid tissues.
关键词
抑制性差减杂交 /
cDNA克隆 /
肌动蛋白结合蛋白 /
淋巴细胞骨架
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Key words
Suppression subtractive hybridization /
cDNA cloning /
Actin associated protein /
Lymphocyte cytoskeleton
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