水稻硅转运子基因的克隆与表达载体的构建

提取水稻根中总RNA,根据NCBI中水稻硅转运子基因Lsil(AB222272)、Lsi2(AB222273)的mR-NA序列设计2对引物,采用RT-PCR技术扩增硅转运子基因Lsil、Lsi2.并利用DNA重组技术分别构建了两个植物表达载体pCAMBIA2300-35S-Lsil-OCS和pCAMBIA2300-35S-Lsi2-OCS.克隆的水稻硅转运子基因Lsil、Lsi2与NCBI中已发表的核苷酸序列的同源性为99.33%、99.65%,氨基酸同源性都为100%.为后续培育其他硅转基因树木或农作物,增加其硅元素转运能力、提高其抗逆性奠定了实验基础.

Cloning and Construction of Expression Vector of Si Transporter Gene in Rice

Total RNA was extracted from tender roots of rice, two pairs of primers were designed according to mRNA sequences of Si transpo rtergenes Lsi1 (AB222272) and Lsi2 (AB222273) on NCB I, Lsi1 and Lsi2 were amplified by these two pairs of primers with RT-PCR.Two plant expression vectors pCAMBIA2300-35S-Lsi1-OCS and pCAMBIA2300-35S-Lsi2-OCS were constructed by DNA recombination technique.The nucleotide sequencing analysis indicated that the cloned fragments showed 99.33%, 99.65% identity to the published sequences, and the amino acid sequences homolog both were 100%.So the proceeding of application on transgene forest trees and crops by the genes Lsi1, Lsi2 will improve efficiency of Si absorption and stress resistance.