大豆类受体蛋白激酶基因rlpk2启动子克隆及初步分析

用双链接头介导的基因组DNA步行技术获得了大豆中与衰老相关的类受体蛋白激酶基因rlpk2 5’侧翼1801bp启动子序列（Prlpk2，GenBank登录号：AY870314）。序列分析结果表明，这一启动子片段中有响应细胞分裂素、脱落酸、赤霉素和生长素等激素作用和响应干旱、寒冷及伤害等逆境胁迫的多个顺式作用元件。构建启动子Prlpk2与报告基因GUS融合基因的双元表达载体pRlpk2．GUS，并通过农杆菌介导法转化大豆幼苗和微型番茄Micro．Tom的子叶外植体，染色结果表明这1801bp的启动子Prlpk2可以有效地驱动GUS基因在大豆幼苗生长点和番茄愈伤组织中瞬时表达。

Cloning and Preliminary Analysis of Promoter of Soybean Receptor-like Protein Kinase Gene rlpk2

5'-Flanking region of rlpk2 gene was cloned by using PCR-based genomic DNA walk- ing method(Fig.1),and designated Prlpk2 (GenBank Accession Number: AY870314). Results of se- quencing analysis showed that Prlpk2 contains sev- eral putative cis-elements that respond to CTK, ABA, GA orIAA, as well as elements that respond to different environmental stresses, such as dehydration, coldness, wounding and etc. An ex- pression vector containing the Prlpk2-GUS fusion gene was constructed (Fig.2) and transferred into soybean seedlings and Micro-Tom cotyledon ex-plants by Agrobacterium-mediated method. A strong transientexpression of GUS was observed in both soybean plumule and Micro-Tom cotyle- don callus (Fig.3), thus confirming the promoter activity of Prlpk2.