雨生红球藻β-胡萝卜素酮化酶基因的cDNA和基因组DNA克隆及序列分析

根据已知的β-胡萝卜素酮化酶(β-carotene ketolase,BKT)的cDNA序列设计引物,利用长距离PCR扩增获得了bkt基因的cDNA完整编码片段及基因组DNA片段,并对这些片断进行了序列测定。cDNA和基因组DNA比对结果表明该基因至少包括5个内含子,它们的剪切位点皆符合GU-AG规律。在比对结果中同时还发现一个19bp的短序列重复,该重复序列在bkt的cDNA和基因组DNA上都发生了多次重复,推测在BKT的表达调控中可能具有一定功能。另外,在序列比对过程中还发现本实验所获得的cDNA和基因组DNA序列与前人报道的cDNA序列之间都存在多处单碱基差异和19bp的短序列差异。

Cloning and sequence analysis of β-carotenoid ketolase cDNA and genomic DNA in Haematococcus pluvialis

This paper had designed primers according to the cDNA sequence reported by Kajiwara,cloned the cDNA and genomic DNA fragments by LD-PCR,and obtained their entire sequences by sequencing.The(alignment) in between cDNA,genomic DNA and cDNA reported by Kajiwara shows that this gene includes five introns at least and all the exon-intron junctions conform to GU-AG spicing rule.Through alignment a 19bp long short repeat unit was found,which exists in cDNA and DNA of bkt for several times.They may have some functions in BKT expression regulation.The diversities from single nucleotides and 19bp short sequence between cDNA sequence reported by Kajiwara and DNA sequences obtained in this paper suggested they have different origins.