Prokaryotic Expressing and Functional Analysis of C3HC4-type Zinc Finger Protein in Nicotiana benthamiana

Abstract

Abstract: C3HC4-type RING finger genes comprise a large family in the plant kingdom and play important roles in various physiologic processes of plant life. In this study, a zinc finger gene was isolated from Nicotiana benthamiana. Sequence analysis indicated that the gene encodes a 24 kD protein with 203 amino acids and contains one typical C3HC4 zinc finger domain and named NbZFP1. The entire coding region of NbZFP1 gene was then cloned into the bacterial expression vectors pGEX-6P-2 and pMAL-C2X. After IPTG induction, the recombinant protein was analyzed by SDS-PAGE, which showed that the expected protein was expressed at a high level in bacterial cells. NbZFP1 gene was cloned into the over-expression vector pBI121, and then transfered into Nicotiana benthamiana through Agrobacterium tumefacien-mediated transformation. According to PCR and Southern blot detection, NbZFP1 gene was successfully integrated into Nicotiana benthamiana genome. Regeneration plants showed type compactness, internode shorten and stem sturdy, also enhanced disease resistance against tobacco mosaic virus（TMV）.

Key words: Zinc finger protein, Recombinant expression, Transgenic tobacco, Disease resistance

Wu Wenxian, Yang Xiufen, Liu Yong, Qiu Dewen. Prokaryotic Expressing and Functional Analysis of C3HC4-type Zinc Finger Protein in Nicotiana benthamiana[J]. Biotechnology Bulletin, 2014, 0(7): 100-105.

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