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选用对虾白斑综合征病毒(White Spot Syndrome Virus,WSSV)囊膜蛋白VP37的功能区段,利用大肠杆菌密码子偏爱性,在氨基酸不变的情况下将VP37中的密码子通过人工合成改为大肠杆菌偏爱型密码子,并克隆至表达载体pBAD/gIIIA中,构成重组载体pBAD/gIIIA-VP37p′。将重组载体转化入大肠杆菌Top10中,在相同条件下用L-阿拉伯糖与未优化的重组菌株Top10-pBAD/gIIIA-VP37p一同诱导。结果显示,与野生型基因VP37p相比,经密码子优化的VP37p′基因表达目的蛋白量占总蛋白的40.5%,明显高于野生型6.5%的目的蛋白表达量。
The functional segment of VP37 of the White Spot Syndrome Virus (WSSV) envelope protein was selected and the codons in VP37 were changed into the large intestine by using the codon preference of E. coli without changing the amino acid Bacillus preferred codon and cloned into the expression vector pBAD / gIIIA to form the recombinant vector pBAD / gIIIA-VP37p ’. The recombinant vector was transformed into E. coli Top10 and induced with L-arabinose under the same conditions with the unoptimized recombinant strain Top10-pBAD / gIIIA-VP37p. The results showed that compared with the wild-type gene VP37p, the codon-optimized VP37p ’gene expressed 40.5% of the total protein, which was significantly higher than that of the wild-type 6.5% of the target protein.