茶多酚对大黄鱼肌肉5个基因片段mRNA稳定性的影响  

Effect of Tea Polyphenols on mRNA Stability of cDNA Fragments of Five Genein Pseudosciaena Crocea Muscle

作　　者：赵进 姚福军[2] 励建荣 刘军 管峰 葛建 戴贤君 Zhao Jin;Yao Fujun;Li Jianrong;Liu Jun;Guan Feng;Ge Jian;Dai Xianjun(College of Life Science,China Jiliang University,Hangzhou 310018;Tonglu County Agricultural Technology Extension Center,Hangzhou 311502;College of Food Science and Engineering,Bohai University,Jinzhou 121013,Liaoning)

机构地区：[1]中国计量大学生命科学学院,杭州310018 [2]桐庐县农业技术推广中心,杭州311502 [3]渤海大学食品科学与工程学院,辽宁锦州121013

出　　处：《中国食品学报》2021年第9期214-224,共11页Journal of Chinese Institute Of Food Science and Technology

基　　金：国家自然科学基金项目(31071514);浙江省重点研发计划项目(2020C02045)。

摘　　要：成功克隆并测序获得大黄鱼肌肉钙蛋白酶基因(Calpain-1、Calpain-2)、钙蛋白酶抑制酶基因(Calpastatin)和蛋白酶家族基因(MuRF-1、MuRF-2)等5个基因片段cDNA,提交Genbank数据库分别获得序列号及长度为:Calpain-1(GeneBank no.KF527412,506bp)、Calpain-2(GeneBank no.KF527413,425bp)、Calpastatin(GeneBank no.KF527409,724bp)、MuRF-1(GeneBank no.KF527410,401bp)和MuRF-2(GeneBank no.KF527411,474bp)。大黄鱼5个基因cDNA片段序列BLAST分析表明:Calpain-1与罗非鱼的同源性达89%,Calpain-2与庸鲽同源性达89%,Calpastatin与鲈鱼的同源性达94%,MuRF-1与大西洋鲑鱼同源性达82%,MuRF-2与罗非鱼同源性达84%。实时定量分析大黄鱼宰杀当天和冷藏期间肌肉中5个基因mRNA的稳定性,得到Calpain-1、Calpastatin、MuRF-1和MuRF-2基因mRNA具有相对稳定性,随着贮藏时间的延长,mRNA水平呈下降趋势,与贮藏时间呈显著负相关性。茶多酚(TP)对冷藏大黄鱼片的保鲜作用,部分原因是通过TP影响大黄鱼片相应mRNA稳定性,进而有效控制冷藏期间鱼肉的新鲜度。The cDNA fragments of Calpain-1,Calpain-2,Calpastatins,MuRF-1 and MuRF-2 in Pseudosciaena crocea were amplified and quantified by RACE-PCR and RT-PCR.Some partial coding sequence for large yellow croaker's five genes was determined and deposited in GenBank,we obtained the accessions and partial coding sequences as follow:Calpain-1(506 bp,no.KF527412),Calpain-2(425 bp,no.KF527413),Calpastatin(724 bp,no.KF527409),MuRF-1(401 bp,no.KF527410)and MuRF-2(474 bp,no.KF527411),respectively.The analytical results of BLAST showed that the partial nucleotide sequence of Calpain-1 cDNA(506 bp)has 89%similarity with Oreochromis niloticus Calpain-1(XM_003441017).The partial cDNA sequence for Calpain-2 is 425 bps long,and the sequence is also closely related to the known Calpain-2 from other species as it shares 89%similarity with Hippoglossus hippoglossus Calpain-2(GQ327966).The nucleotide sequence of Calpastatin consists of 724 bps,and it displayed high sequence homology with the previously reported Calpastatin in other species with 94%similarity with Dicentrarchus labrax(FQ310506).The nucleotide sequence of MuRF-1 cDNA is 401 bps in length and the sequence shows 82%similarity with Salmo salar(NM_001279046).The cDNA sequence for MuRF-2 is 474 bps,and it shares 84%similarity with Oreochromis niloticus(XM_003458136).In our study,the four gene-specific mRNA levels had either a longer post-mortem interval or a slower decreasing speed in the TP samples than in those in the control samples,except that no differences were observed in the mRNA level of the Calpain-2 gene when assessed with respect to the post-mortem storage time.The preservation effect of tea polyphenols(TP)on large yellow croaker fillets is partly due to the effect of TP on the corresponding mRNA stability fillets,thus effectively controlling the freshness of fish during refrigeration.

关 键 词：冷藏 大黄鱼 蛋白酶基因 MRNA稳定性 新鲜度 

分 类 号：TS254.1[轻工技术与工程—水产品加工及贮藏工程]