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从鳗草种中克隆 TPS 基因及其在水稻中的遗传转化功能鉴定。

Cloning of TPS gene from eelgrass species Zostera marina and its functional identification by genetic transformation in rice.

机构信息

Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.

出版信息

Gene. 2013 Dec 1;531(2):205-11. doi: 10.1016/j.gene.2013.09.016. Epub 2013 Sep 11.

Abstract

The full-length cDNA sequence (2613 bp) of the trehalose-6-phosphate synthase (TPS) gene of eelgrass Zostera marina (ZmTPS) was identified and cloned. Z. marina is a kind of seed-plant growing in sea water during its whole life history. The open reading frame (ORF) region of ZmTPS gene encodes a protein of 870 amino acid residues and a stop codon. The corresponding genomic DNA sequence is 3770 bp in length, which contains 3 exons and 2 introns. The ZmTPS gene was transformed into rice variety ZH11 via Agrobacterium-mediated transformation method. After antibiotic screening, molecular characterization, salt-tolerance and trehalose content determinations, two transgenic lines resistant to 150 mM NaCL solutions were screened. Our study results indicated that the ZmTPS gene was integrated into the genomic DNA of the two transgenic rice lines and could be expressed well. Moreover, the detection of the transformed ZmTPS gene in the progenies of the two transgenic lines was performed from T1 to T4 generations; and results suggested that the transformed ZmTPS gene can be transmitted from parent to the progeny in transgenic rice.

摘要

我们鉴定并克隆了鳗草(Zostera marina)海藻糖-6-磷酸合酶(TPS)基因的全长 cDNA 序列(2613 bp)。鳗草是一种终生在海水中生长的种子植物。ZmTPS 基因的开放阅读框(ORF)区域编码一个 870 个氨基酸残基的蛋白质和一个终止密码子。相应的基因组 DNA 序列长 3770 bp,包含 3 个外显子和 2 个内含子。ZmTPS 基因通过农杆菌介导的转化方法被转化到水稻品种 ZH11 中。经过抗生素筛选、分子特征分析、耐盐性和海藻糖含量测定,筛选出了两株能耐受 150 mM NaCl 溶液的转基因株系。我们的研究结果表明,ZmTPS 基因已整合到两个转基因水稻株系的基因组 DNA 中,并能良好表达。此外,还对两个转基因株系的后代进行了转化的 ZmTPS 基因的检测,检测范围从 T1 代到 T4 代;结果表明,转化的 ZmTPS 基因可以在转基因水稻中从亲代传递给后代。

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