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应用蛋白质组学研究低温胁迫下龙眼叶片蛋白质组变化时发现碳酸酐酶(CA)蛋白下调表达。利用RT-PCR方法克隆CA基因的全长cDNA,GenBank登录号JN033201,长度为1119bp,包括1个966bp的开放阅读框,编码321bp的氨基酸序列,同源性分析表明,12个不同植物同源性为81%～88%。龙眼CA基因具有典型的CA结构域,并且非常保守。实时荧光定量分析结果表明,CA在龙眼根、茎、叶中都有表达,为组成型表达,在叶中的表达量最高,茎和根中的表达量最少。CA基因在低温胁迫下随着低温胁迫时间的延长而发生变化。将CA在大肠杆菌中表达,获得1个约40.5kD的外源蛋白。推测CA表达与低温胁迫有关。
Using proteomics, the expression of carbonic anhydrase (CA) protein was down-regulated when the proteome was changed under low temperature stress. The full-length cDNA of CA gene was cloned by RT-PCR. GenBank accession number JN033201 was 1119bp in length, including a 966bp open reading frame (ORF) encoding a 321bp amino acid sequence. Homology analysis showed that 12 different plant homologies 81% ~ 88%. The longan CA gene has a typical CA domain and is very conserved. The results of real-time fluorescence quantitative analysis showed that CA was expressed in roots, stems and leaves of longan, and was constitutively expressed in leaves, with the highest expression in leaves and the least expression in stems and roots. CA gene changed with the extension of chilling stress time under low temperature stress. CA was expressed in E. coli to obtain a foreign protein of about 40.5 kD. It is speculated that CA expression is related to low temperature stress.