Construction and Transformation of Borago officinalis BoD6D Gene Vector

doi:10.13560/j.cnki.biotech.bull.1985.2020-0791

Abstract

Abstract:

The purpose of this study is to lay a foundation for increasing the content of γ-linolenic acid in oil crops via constructing Borago officinalis BoD6D transforming vector by means of plant genetic engineering. The BoD6D gene was cloned using borage genomic DNA as a template. Then,a yeast expression vector was constructed and transformed into Saccharomyces cerevisiae. The fatty acids were extracted to have methyl esterification reaction,and the fatty acid content was analyzed by gas chromatography. Concurrently,the plant binary expression vector was constructed,and the Arabidopsis thaliana was transformed by the dipping method mediated by Agrobacterium,and finally the transgenic Arabidopsis was identified by resistance screening and PCR. Analysis found that the coding region of BoD6D gene did not contain introns and could be directly used for subsequent functional research. The expression vector pYES2-BoD6D of S. cerevisiae was successfully constructed,and the results of gas chromatography showed that BoD6D gene successfully catalyzed linoleic acid to γ-linolenic acid. The plant binary expression vector pBI121-BoD6D was successfully constructed. The results of kanamycin resistance screening and PCR identification demonstrated that the Arabidopsis transformed plants with the BoD6D gene were ultimately obtained. The above results indicate that the borage genomic DNA can be directly used as a template to easily obtain functional BoD6D gene for transgenic plant research.

Key words: Borago officinalis, BoD6D gene, γ-linolenic acid, yeast expression vector, plant expression vector

GONG Yuan-yong, ZHAO Li-hua, YAN Fei, GUO Shu-qiao, SHU Hong-mei, NI Wan-chao. Construction and Transformation of Borago officinalis BoD6D Gene Vector[J]. Biotechnology Bulletin, 2021, 37(3): 227-232.

Figures/Tables 6

References 17

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