烟草硼外向转运体NIP5;1的克隆和表达分析

为研究烟草中的硼转运机制,克隆了烟草硼外向转运体基因,并分析其表达特点。采用生物信息学的方法预测烟草硼外向转运体的开放阅读框,并通过PCR的方式克隆后测序验证；其相应蛋白质的结构特点亦用生物信息学手段分析。该基因在烟草不同组织和对不同处理响应的表达差异采用半定量RT-PCR的方法检测。结果显示克隆到烟草硼外向转运体NtNIP5;1的开放阅读框,全长894 bp,编码297个氨基酸,具有MIP家族典型的结构域HLNPSLTIA,GenBank登录号为KF611892。其蛋白质的二级结构以α-螺旋为主,有5个跨膜结构域。RT-PCR结果发现,NtNIP5;1在根和叶中均有表达,但在叶中表达水平较高； 6-BA、SA和ABA均可抑制其表达。

Cloning and Expression Analysis of Boron influxTransporter NIP5;1 from Nicotiana tabacum

To understand the mechanism of boron transportation in tobacco, the boron influx transporter gene was cloned in Nicotiana tabaccum, and its expression was analyzed. The open reading frame (ORF) of boron influx transporter was predicted by biology informatics methods, and cloned by PCR technique, verified by sequencing. Its structure was also predicted by biology informatics methods. Semi-quantitative RT-PCR was used to monitor its expression levels in different tissues of tobacco with different treatments. Results showed that the complete ORF sequence of tobacco influx boron transporter, NtNIP5;1 was cloned. Its ORF contains 894 bp, encodes 297 amino acids and includes MIP protein signature sequence HLNPSLTIA. GenBank accession number is KF611892. Its protein consists of α-helix mainly in secondary structure, and was composed of five transmembrane domains. Results of RT-PCR showed that NtNIP5;1 were expressed in roots and leaves, and showed higher levels in leaves. Its expression was inhibited by 6-BA, SA and ABA.