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Cloning of Dehydrin Gene from Ammopiptanthus Mongolicus and It’s Genetic Transformation into Tobacco

Author: QiaoHuiLei
Tutor: WangRuiGang；GuoJiuFeng
School: Inner Mongolia Agricultural University
Course: Biochemistry and Molecular Biology
Keywords: Dehydration gene Clone Plant expression vector Genetic transformation
CLC: Q943.2
Type: Master's thesis
Year: 2009
Downloads: 103
Quote: 2
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Abstract


In this paper, the dehydration gene in drought-resistant Ammopiptanthus mongolicus was successfully cloned. The gene was transformed into tobacco by Agrobacterium tumefaciens, and the transgenic plants were identified by PCR. These transgenic plants will be helpful in further studying the dehydration gene. The major results are as the following:1. Gene specific rimers were designed according to the dehydration gene’s sequence, and the gene was cloned from Ammopiptanthus mongolicus using the genomic DNA as template. The full length gene was 1106bp without any intron, including 98bp of 5’UTR and 456bp of 3’UTR, it encoded a polypeptide of 183 amino acids.2. The target fragment was amplified through specific primers Containing BstEⅡand BglⅡrestriction endonuclease sites, and was inserted into the GUS expression region of binary vector pCAMBIA3301-DHN. The plant expression construct was transformed into Agrobacterium tumefaciems by freeze-thaw method. Colony PCR detection showed that the dehydration gene plant expression recombinat plasmid was successfully constructed and transformed into Agrobacterium.3. The binary vector carrying the dehydration gene was transformed into the sterilized tobacco leaves to obtain transgenic plants. PCR detection with the recombinant expression vector as positive control and the non-transgenic tobacco as negative control was performed using the gene specific primers. The results revealed that the dehydration gene was successfully transferred into tobacco.

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CLC: > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering
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