多浆旱生植物霸王液泡膜H+-PPase基因片段的克隆及序列分析

摘要

Degenerate primers were designed based on conserved sequences of H+-Pase genes from other plants.Total RNA was extracted from the leaves of Zygophyllum xanthoxylum and the H+-PPase gene fragment was obtained by reverse transcription polymerase chain reaction (RT-PCR). It was cloned into PUCm-T vector and the positive clones identified by PCR were sequenced. Three H+-PPase gene fragments were gained from Z.xanthoxylum, each of which contains 898 bp and encodes a peptide of 299 amino acids. Similarity comparisons show that they share over 69％ similarity in nucleotide sequence and over 78％ similarity in amino acid sequence with those of other plant H+-PPase genes in GenBank.%根据其他植物H+-PPase基因的保守序列设计-对简并性引物,以霸王叶片总RNA为模板,采用RT-PCR的方法扩增出H+-PPase基因片段并克隆到PUCm-T载体.阳性克隆经PCR鉴定后进行测序,序列分析结果表明,克隆到3个同源的序列片段(898 bp),编码299个氨基酸;所得序列与GenBank中注册的高等植物H+-PPase 基因核苷酸序列的同源性均在69%以上、氨基酸序列的同源性达78%以上.