枇杷β-胡萝卜素羟化酶基因片段的克隆和序列分析

Molecular cloning and sequence analysis of β-carotene hydroxylase gene fragment from loquat

根据蔷薇科植物β-胡萝卜素羟化酶基因的保守区序列,设计1对PCR引物,以枇杷基因组DNA为模板,采用PCR方法扩增出长约450 bp的DNA片段,克隆人pMD-18T载体中,测得该基因片段长为440 bp,编码145个氨基酸,属于开放阅读框的一部分,不含内含子。在GenBank中进行同源性检索,结果该序列与苹果β-胡萝卜素羟化酶基因同源性为95%,证明它属于枇杷的β-胡萝卜素羟化酶基因。

A pair of primer, designed from conserved regions of β-carotene hydroxylase from rosaceae, was used to amplify homogeneous gene by polymerase chain reaction （PCR）. As a result, one about 450 bp fragment was obtained, and then cloned into pMD-18T vector and sequenced. The result suggested that the 440 bp belong to a part of ORF （open reading frame） and encode 145 amino acids. It is suggested that the sequence is 95% identical to the β-carotene hydroxylase gene from apple, it could be deduced as β-carotene hydroxylase gene fragment of loquat.