Molecular cloning of AGPase large subunit I gene and construction of its sense, antisense and RNAi expression vectors

To study the function of AGPase large subunit I gene (LSU I) in synthesis of starch in wheat plants, full cDNA sequences (1947 bp) of a LSU I gene (GenBank No. DQ839506) were cloned from grains of common wheat (Triticum aestivum, Yujiao 2 cultivar) with RT-PCR, and its sense, antisense and RNAi vect...

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Bibliographic Details
Main Authors: KANG Guo-zhang, ZHANG Meng-qin, GUAN Chun-yun, GUO Tian-cai, ZHU Yun-ji
Format: Article
Language:English
Published: Zhejiang University Press 2007-11-01
Series:浙江大学学报. 农业与生命科学版
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Online Access:https://www.academax.com/doi/10.3785/1008-9209.2007.06.0607
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Summary:To study the function of AGPase large subunit I gene (LSU I) in synthesis of starch in wheat plants, full cDNA sequences (1947 bp) of a LSU I gene (GenBank No. DQ839506) were cloned from grains of common wheat (Triticum aestivum, Yujiao 2 cultivar) with RT-PCR, and its sense, antisense and RNAi vectors were also constructed. The coding regions of LSU I were 1569 bp (from 117-1685 bp), which shared high homogeneity with Z21969 gene sequences. The cloned LSU I gene had two different bases with the Z21969 (T/C, at 851 bp site; and G/A, at 926 bp site), but the two differences did not change their amino sequences, implying the function of the cloned LSU I was not affected. The sense and antisense expression vectors of LSU I were constructed, in which the coding region of the gene was placed under the 35S promoter in either sense or antisense orientation. In addition, especial 250 bp fragments of LSU I were also cloned, and this fragment was inserted into pFGC5941 vector to construct RNAi expression vector of LSU I. These constructed vectors provided a good background to study the function of LSU I.
ISSN:1008-9209
2097-5155