Cloning and sequence analysis of allophycocyanin genes from Porphyra haitanensis

Abstract

以野生坛紫菜色素体DNA为模板, 通过PCR扩增获得编码坛紫菜别藻蓝蛋白α亚基和β亚基的序列及两者之间的间隔序列,该序列全长为1 055 bp,其中编码α和β亚基的序列均为486 bp,间隔序列为83 bp.该序列与GenBank收录的其他4种红藻相关序列的同源性在75.6%~87.4%,与2种蓝藻的同源性分别为66.9%,68.5%,其中编码区同源性更高.On the basis of the sequences of allophycocyanins from several species in genbank,a pair of degenerate primers were designed and synthesized. A specific fragment about 1 055 bp in size was obtained after PCR amplification using the cpDNA of Porphyra haitanensis as template. Then the purified fragment of DNA was cloned into vector pMD18-T, and sequenced after the identification of the recombinants using PCR and endonuclease digest. The results indicate that α and β subunit genes of allophycocyanin are obtained from Porphyra haitanensis. The fragment size is (1 055 bp), including α and β subunits coding regions (486 bp) each and the spacer between them is 83 bp.The genes were submitted to genbank and their genbank accession number was AY372218.福建省重大农业科技资助项目(2001Z017);; 国家海洋“863”资助项目(2002AA603023

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