食品与机械2025，Vol.41Issue(11)：1-8,8.DOI:10.13652/j.spjx.1003.5788.2025.80437
凡纳滨对虾原肌球蛋白的异源重组表达
Heterologous recombinant expression of tropomyosin from Litopenaeus vannamei
摘要
Abstract
[Objective]To obtain recombinant tropomyosin(TM).[Methods]In this study,the gene sequence of a TM homologous protein is obtained from the National Center of Biotechnology Information(NCBI)database,and specific primers are designed accordingly.Subsequently,using the cDNA of Litopenaeus vannamei meat as the template,the TM coding gene sequence of L.vannamei is amplified by PCR and sequenced.Finally,the prokaryotic recombinant expression system for L.vannamei TM is established using the Escherichia coli heterologous recombinant expression vector pET29a.[Results]After agarose gel electrophoresis(AGE)of the RNA samples,the band is clear and bright,with no diffusion observed in the lanes.The cDNA sample displays a clear band around 300 bp,indicating successful extraction of the total RNA samples from L.vannamei meat with intact structures and subsequent reverse transcription into cDNA.The PCR results indicate a single band at 900 bp for the TM coding gene of L.vannamei.BLAST analysis of the gene sequence shows that the TM coding gene is highly homologous(99.77%)to the known TM coding gene of L.vannamei.This study further constructs the TM recombinant expression vector pET29a-TM.SDS-PAGE analysis confirms efficient expression of the target protein in the host strain,yielding a soluble recombinant TM protein band with a relative molecular weight of approximately 3.7×104.[Conclusion]This study successfully clones the TM coding gene from L.vannamei and constructs its prokaryotic expression system,enabling the efficient production of soluble recombinant TM protein.关键词
原肌球蛋白/食物过敏/凡纳滨对虾/基因克隆/重组表达Key words
tropomyosin/food allergy/Litopenaeus vannamei/gene cloning/recombinant expression引用本文复制引用
李言初,杨玉莹,胡卫成,刘书成,魏帅..凡纳滨对虾原肌球蛋白的异源重组表达[J].食品与机械,2025,41(11):1-8,8.基金项目
国家自然科学基金面上项目(编号:32272245) （编号:32272245）
国家虾蟹产业技术体系(编号:CARS-48) （编号:CARS-48）
