莲雾乙烯受体基因保守序列的克隆及分析

The Cloning and Analysis of the Conserved Sequence of Ethylene Receptor Gene in Wax Apple

用莲雾的叶片提取基因组DNA作为模板,根据乙烯受体基因的保守序列设计引物,进行PCR扩增,得到长度约1 5kb的目的片段,将其克隆到pGEMT easy载体转化大肠杆菌,提取质粒,进行限制性内切酶图谱分析及序列测定。结果表明,该目的片段全长为1400bp,与栽培稻(Oryzasativa)的osers基因组DNA可比对序列的一致性为98 68%。在所应用的12个内切酶中,该片段内部没有切点,根据序列特征和与其他乙烯受体基因序列比较,推测该片段有2个内含子和3个外显子片段组成,外显子总长为972bp,编码324个氨基酸,其中第197～324氨基酸残基与组氨酸蛋白激酶的磷酸基团接受域有同源性。

A ethylene receptor protein gene fragment of about 1.5 kb was obtained by PCR amplifying from the genomic DNA of wax apple (Syzygium samarangense) leaves using a pair of primers designed according to the conserved sequence of ethylene receptor protein gene. This fragment was linked into the pGEM T-easy vector, then transformed the E.coli DH5α. This fragment was analyzed by restriction enzyme maping and sequenced. The result revealed that the length of this fragment was 1400 bp contained two introns and three exons. The whole length of exon was 974 bp, coding 324 amino acids. Sequence compared analysis suggested that this fragment was most similar to the sequence of rice(Oryza sativa) ethylene receptor gene (osers) ,the identity was 98.68%.