The Construction Of Expression Vector Containing DNA Sequence Of A And B Strand Of Human Insulin And Its Genetic Transformation Of Soybean

The theory of plant reactor of medicine was advanced in the end of 1980s. Nowadays,plant reactor have received more and more attention because theoretically, it is a prospective cheaper and safer way to produce medical proteins or polypeptides. A great number of medical proteins or polypeptides were already expressed effectively in plant. But there is a longer way before us before plant reactors are used to produce commercial medical proteins or polypeptides. Here, we synthesized the DNA sequence of A and B strand of human insulin with soybean biased codon after analyzing the soybean codon bias and introduced it into conventional soybean strain. "Ji30" through Agrobacterium-mediated transformation system. Consequently, paved the way for the expression of human insulin gene in soybean. The major results in the paper are abstracted as follows:1 we synthesized the DNA sequence of A and B strand of human insulin with soybean biased codon after analyzing the soybean codon bias according to the cDNA data of soybean in GeneBank.2.Insert the above DNA sequence into the pBI121 with CaMV35S and constructed the soybean binary expression vector pBI 121-IN. Then, we i ntroduced pBH21-IN into the cell of Agrobacterium tumefaciens EHA105 and obtain EHA105 (pBI121-IN) which was positive in PCR detection.3.By optimizing the parameters of co-culture between EHA105 (pBI121-IN) and cotyledonary node of soybean Ji30, we established a effective Agrobacterium EHA105-mediated genetic transformation system of cotyledonary node of Ji30.4. By the established EHA105-mediated genetic transformation system of Ji30, we obtained Km resistant seedling in rooting medium.