Cloning and Transformation of Gene GmGW2 in Soybean

doi:10.13560/j.cnki.biotech.bull.1985.2015.08.012

Abstract

Abstract: GW2 encodes a previously unknown RING-type protein with E3 ubiquitin ligase activity, which is located in cytoplasm. It is known to function in the degradation by the ubiquitin-proteasome pathway. Loss of GW2 function increased cell numbers. It was reported that the mutation of GW2 lead to change the size of rice seeds. According to rice GW2 gene sequence, blast in the NCBI, we found a soybean GW2 gene which is homologous. We cloned the target gene and carried on bioinformatics analysis, then we constructed a over-expression vector pCAMBIA1300∷GmGW2∷EYFP, and transformed into soybean named zhonghuang10 by Agrobacterium-mediated method. Seeds were germinated on sterile moistened paper towels in Petri dishes for 24 h then infected the immature embryo by Agrobacterium EHA105 which obtained recombining vector. When the explants have 3 compound leaves, we extract the genome DNA and PCR identified the report gene EYFP. The content of EYFP is enhanced in PCR positive plant was observed by LSCM. It is demonstrated the transformation of target gene is successful. The sequence analysis showed that the target gene have higher homology with those of Alfalfa and Gram.

Key words: GmGW2, soybean, phylogenetic tree, transformation, EYFP

Xiong Hewen, Wu Zhihui, Duan Siyu, Xie Hao, Qin Yutao, Guo Bei. Cloning and Transformation of Gene GmGW2 in Soybean[J]. Biotechnology Bulletin, 2015, 31(8): 82-87.

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