拒盐型盐生植物小花碱茅肌动蛋白基因片段的克隆及序列分析

Cloning and Sequence Analysis of Actin Gene Fragment from Salt-Exclusion Halophyte Puccinellia tenuiflora

本研究根据其它植物Actin基因的保守序列设计一对简并性引物,以拒盐型盐生植物小花碱茅根部总RNA为模板,采用RT-PCR的方法扩增出Actin基因片段并克隆到PUCm-T载体,阳性克隆经PCR检测后进行测序,在GenBank中注册;序列分析结果表明:该片段长约600bp,编码198个氨基酸;所得序列与GenBank中注册的其它植物Actin基因序列同源性均在84%以上,与其它肌动蛋白的氨基酸序列同源性达94%以上。

In this paper degenerate primers were designed based on the conserved sequences of the Actin gene from other plants. Total RNA was extracted from the roots of Puccinellia tenuiflora. A ctin gene fragment was obtained by reverse transcription polymerase chain reaction （RT-PCR） and cloned into PUCm-T vector. The positive clone identified by PCR was sequenced and registered in GenBank. The sequencing result revealed that the A ctin gene fragment from Puccinellia tenuiflora contains about 600 bp, encoding 198 amino acids. Homology comparison with Actin gene sequences of other plants in the GenBank and with other Actins showed that it shared over 84% nucleotide sequence homology and 94% amino acid sequence homology.