Cloning of Kunitz- type trypsin inhibitor gene SKTI3 and construction of its plant expression vector
CHAI Xiao-jie, LU Pin, ZHANG Yu, WANG Pi-wu
1. School of Life Science and Technology, Dalian Fisheries Univ. , Dalian 116023, China 2. Faculty of Biotechnology, Jilin Agricultural University, Changchun 130118, China
Abstract: The gene SKTI3 of soybean Kunitz - type trypsin inhibitor was cloned by PCR using soybean genomic DNA as template and inserted into pMD18 -T vector. DNA sequence analysis showed that the cloned fragment consisted of 654 bp and shared 99% identity with the SKTI3 sequence published on Genbank. The cloned gene was inserted to the downstream of 35S promoter in the binary vector pBI121 to construct the recombinant plasmid pBISKTI3. Then, pBISKTI3 was mobilized into Agrobacterium tumefaciens strain EHA105 by freeze -melting methods. The plant expression vector containing SKTI3 gene was obtained.
柴晓杰, 吕品, 张宇, 王丕武. 胰蛋白酶抑制剂SKTI3基因的克隆及其植物表达载体的构建[J]. 大连海洋大学学报, 2007, 22(5): 384-386.
CHAI Xiao-jie, LU Pin, ZHANG Yu, WANG Pi-wu. Cloning of Kunitz- type trypsin inhibitor gene SKTI3 and construction of its plant expression vector. Journal of Dalian Ocean University, 2007, 22(5): 384-386.