Construction of Overexpression Vector of CONSTANS Gene Plant in Brassica napus and Production of Transgenic Plants
1 Crop Research Institute, Sichuan Academy of Agricultural Sciences, Chengdu, 610066, China
2 Zoeve Seed, Chengdu, 610066, China
Molecular Plant Breeding, 2018, Vol. 9, No. 10 doi: 10.5376/mpb.2018.09.0010
Received: 13 Sep., 2018 Accepted: 13 Nov., 2018 Published: 14 Dec., 2018
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This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Zheng B.C., Li H.J., Zhang J.F., Cui C., Chai L., Jian J., Wu X.M., and Jiang L.C., 2018, Construction of overexpression vector of CONSTANS gene plant in brassica napus and production of transgenic plants, Molecular Plant Breeding, 9(10): 73-79 (doi: 10.5376/mpb.2018.09.0010)
The CONSTANS target gene from Brassica napus was cloned, and the full length amplified primer of it was designed and amplified on the sequence released on NCBI. The target gene fragments were sequenced and the restriction loci were analyzed. The enzyme primers (BamHⅠ, SacⅠ) were designed, and the target gene fragments were connected to the eukaryotic expression vector pBI121 through BamHⅠ and SacⅠ double enzyme digestion. PCR identification result indicated that PBI121+CONSTANS overexpression vector was successfully constructed. The overexpression vector was transformed into the competent EHA105 cell, and the seeds of T0 were infected by Arabidopsis thaliana. The positive plants were screened by MS solid medium containing Cara (50 mg/L). PCR identification showed that PBI121+CONSTANS overexpression vector was successfully introduced into Arabidopsis thaliana and 12 transgenic positive seedlings were obtained, which did the preparation for the gene function analysis.
Brassica napus; Overexpression vector; Positive plants