杜氏盐藻2种碳酸酐酶基因跨内含子DNA序列的克隆和鉴定  

Cloning and identification of the cross-intron genomic DNA sequences of the two carbonic anhydrase genes from Dunaliella salina

作　　者：吕玉民[1] 姜国忠[1] 牛向丽[1] 谢华[1] 薛乐勋[1] 

机构地区：[1]郑州大学细胞生物学研究室,郑州450052

出　　处：《郑州大学学报（医学版）》2004年第1期23-25,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家高技术研究发展 ( 863 )计划 2 0 0 2AA62 80 5 0 ;河南省重大科技攻关基金资助项目 0 12 2 0 3 2 5 0 0 ;河南省杰出人才创新基金资助项目 0 2 2 10 0 190 0

摘　　要：目的 :克隆杜氏盐藻双拷贝碳酸酐酶 (DCA1 )和碳酸酐酶 (CA1 )基因跨内含子基因组DNA序列。方法 :利用跨内含子PCR方法 ,从杜氏盐藻基因组中克隆DCA1和CA1基因跨内含子基因组DNA序列。结果 :DCA1基因跨内含子长度为 4 4 0 7bp ,含 7个外显子 ,由此推定的氨基酸序列长度为 5 5 5个氨基酸残基 ;而CA1基因跨内含子长度为 4 4 73bp ,含 8个外显子 ,其推定的氨基酸序列长度为 4 98个氨基酸残基。这 2种基因的所有外显子 -内含子交接点处序列都遵守GT -AG规律。结论Aim: To clone the cross-intron genomic DNAs of the duplicated carbonic anhydrase (DCA1) and carbonic anhydrase (CA1) genes from Dunaliella salina. Method: The sense and antisense primers of a cross-intron PCR were designed according to the 5' - and 3' -UTR sequences of DCA1 and CA1 cDNAs. The cross-intron PCR amplification was performed with Dunaliella salina genome DNA as a template. Results: The cross-intron genomic DNA sequence of DCA1 gene had 4 407 bases which contained seven exons and six introns, and the deduced amino acid sequence of the DCA1 coding region had 555 amino acids. The cross-intron genomic DNA sequence of CA1 gene had 4 473 bases in which there were eight exons and seven introns, and the deduced amino acid sequence of the CA1 coding region had 498 amino acids. Splicing junction sequences of all the exon-introns conformed to the GT-AG rule in the cross-intron genomic DNA sequences of DCA1 and CA1 genes. Conclusion: The sequences obtained by cross-intron PCR are identified to be cross-intron genomic DNA sequences of DCA1 and CA1 genes of Dunaliella salina, respectively.

关 键 词：杜氏盐藻 双拷贝碳酸酐酶 碳酸酐酶 跨内含子基因组DNA 

分 类 号：Q781[生物学—分子生物学]