[1]李武峰,赵婧微,孙瑜彤,等.广灵驴CAST基因的克隆与表达[J].福建农林大学学报(自然科学版),2020,49(05):652-659.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2020.05.012]
 LI Wufeng,ZHAO Jingwei,SUN Yutong,et al.Cloning and expression analysis of CAST gene in Guangling donkey[J].,2020,49(05):652-659.[doi:10.13323/j.cnki.j.fafu(nat.sci.).2020.05.012]
点击复制

广灵驴CAST基因的克隆与表达()
分享到:

福建农林大学学报(自然科学版)[ISSN:1671-5470/CN:35-1255/S]

卷:
49卷
期数:
2020年05期
页码:
652-659
栏目:
动物科学
出版日期:
2020-09-18

文章信息/Info

Title:
Cloning and expression analysis of CAST gene in Guangling donkey
文章编号:
1671-5470(2020)05-0652-08
作者:
李武峰1 赵婧微1 孙瑜彤1 杜 敏2
1.山西农业大学生命科学学院,山西 太谷 030801; 2.美国华盛顿州立大学动物科学系,美国华盛顿州普尔曼 99164-6310
Author(s):
LI Wufeng1 ZHAO Jingwei1 SUN Yutong1 DU Min2
1.College of Life Sciences, Shanxi Agricultural University, Taigu, Shanxi 030801, China; 2.Department of Animal Sciences, Washington State University, Pullman, WA 99164-6310, USA
关键词:
广灵驴 钙蛋白酶抑制(CAST)基因 基因克隆 组织表达
Keywords:
Guangling donkey calpastatin(CAST)gene clone tissue expression
分类号:
S822
DOI:
10.13323/j.cnki.j.fafu(nat.sci.).2020.05.012
文献标志码:
A
摘要:
对广灵驴钙蛋白酶抑制(CAST)基因进行克隆及序列分析,并用实时荧光定量PCR技术对其在心、肝、脾、肺、肾和背最长肌等6种组织中的表达水平进行分析,探究CAST基因对驴肉品质的影响.结果表明,广灵驴CAST基因的CDS编码区全长2 427 bp,共编码808个氨基酸,将测序序列提交至NCBI,得到的GenBank登录号为MN231002.广灵驴CAST蛋白的氨基酸序列与马、牛、人、猪、山羊、绵羊、小鼠、鸡的同源性分别为98.6%、77.1%、70.8%、77.5%、70.1%、60.9%、65.1%和41.5%; CAST基因在广灵驴6种组织中均存在表达,其中,表达量最高的组织为背最长肌.本试验成功获得了广灵驴的CAST基因,并发现其在背最长肌中高度表达,为进一步研究CAST基因在肌肉嫩度方面的表达调控功能提供参考.
Abstract:
In order to explore the effect of calpastatin(CAST)gene on meat quality of Guangling donkey, the CAST gene was cloned and sequenced, and the expressional levels of CAST gene in different tissues of heart, liver, spleen, lung, kidney and longissimus dorsi muscle were analyzed by real-time PCR. The results showed that the CAST gene fragment included 2 427 bp, encoded 808 amino acids, whose accession number is MN231002 in GenBank. The amino acid sequence of Guangling donkey CAST protein was 98.6%, 77.1%, 70.8%, 77.5%, 70.1%, 60.9%, 65.1% and 41.5% homologous to horse, cattle, human, swine, goat, sheep, mouse and chicken. The CAST gene was expressed in all 6 tissues, whose expression level was maximized in longissimus dorsi and followed by heart. The study provides theoretical basis for further study of the expression and regulatory function of CAST gene in muscle tenderness.

参考文献/References:

[1] HANNA R A, CAMPBELL R L, DAVIES P L. Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin[J]. Nature, 2008,456:409-412.
[2] 左北瑶,钱宏光.CAST基因研究进展[J].中国草食动物科学,2011,31(6):64-68.
[3] ILIAN M A, MORTON J D, KENT M P, et al. Intermuscular variation in tenderness: association with the ubiquitous and muscle-specific calpains[J]. Journal of Animal Science, 2001,79(1):122-132.
[4] 李婧,郭月英,任霆,等.钙蛋白酶抑制蛋白及CAST基因对肉品质及嫩度的影响[J].食品工业,2013(6):141-144.
[5] KOOHMARAIE M, SEIDEMANN S C, SCHOLLMEYER J E, et al. Effect of post-mortem storage on Ca++-dependent proteases, their inhibitor and myofibril fragmentation[J]. Meat Science, 1987,19(3):187-196.
[6] DELGADO E F, GEESINK G H, MARCHELLO J A, et al. The calpain system in three muscles of normal and callipyge sheep[J]. Journal of Animal Science, 2001,79(2):398-412.
[7] 燕凤.绵羊CAPN1、CAST基因mRNA的发育性表达规律及其对肉品质的影响[D].杨凌:西北农林科技大学,2008.
[8] 孙立彬,孟和,李婧,等.猪钙蛋白酶抑制蛋白基因PCR-RFLP多态性与肉质性状及背膘厚间的关系分析[J].上海交通大学学报(农业科学版),2005,23(1):57-62.
[9] 周楠,韩国才,柴晓峰,等.驴的产肉、理化指标及加工特性比较研究[J].畜牧兽医学报,2015,46(12):2314-2321.
[10] SIMMS D, CHOMCZYNSK P. TRIzolTM: A new reagent for optimal single-step isolation of RNA[J]. Focus, 1993,15(4):99-102.
[11] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-△△CT method[J]. Methods, 2001,25(4):402-408.
[12] GUROFF G. A neutral, calcium-activated proteinase from the soluble fraction of rat brain[J]. Journal of Biological Chemistry, 1964,239(1):149-155.
[13] MURAHI T. Intracellular regulatory system involving calpain and calpastatin[J]. Biochemistry International, 1989,18(2):263-294.
[14] EMORI Y, KAWASAKI H, IMAJOH S, et al. Endogenous inhibitor for calcium-dependent cysteine protease contains four internal repeats that could be responsible for its multiple reactive sites[J]. Proceedings of the National Academy of Sciences, 1987,84(11):3590-3594.
[15] TAKANO E, MAKI M, MORI H, et al. Pig heart calpastatin: identification of repetitive domain structures and anomalous behavior in polyacrylamide gel electrophoresis[J]. Biochemistry, 1988,27(6):1964-1972.
[16] LEE W J, HATANAKA M, MAKI M. Multiple forms of rat calpastatin cDNA in the coding region of functionally unknown amino-terminal domain[J]. Biochimica et Biophysica Acta(BBA)-Gene Structure and Expression, 1992,1129(2):251-253.
[17] KILLEFER J, KOOHMARAIE M. Bovine skeletal muscle calpastatin: cloning, sequence analysis, and steady-state mRNA expression[J]. Journal of Animal Science, 1994,72(3):606-614.
[18] GOLL D E, THOMPSON V F, HONGQI L I, et al. The calpain system[J]. Physiological Reviews, 2003,83(3):731-801.
[19] OTSUKA Y, GOLL D E. Purification of the Ca2+ dependent proteinase inhibitor from bovine cardiac muscle and its interaction with the millimolar Ca2+-dependent proteinase[J]. Journal of Biological Chemistry, 1987,262(12):5839-5851.
[20] 邱建丁,梁汝萍,谭学才,等.膜蛋白跨膜区段的预测分析[J].高等学校化学学报,2004,25(5):831-836.
[21] UEMORI T, SHIMOJO T, ASADA K, et al. Characterization of a functional domain of human calpastatin[J]. Biochemical and Biophysical Research Communications, 1990,166(3):1485-1493.
[22] 李静.CAST基因及钙蛋白酶系统的研究进展[J].安徽农业科学,2007(15):4618-4619.
[23] 王晓龙,王秀然,卢天成,等.蛋白质糖基化修饰的研究进展[J].基因组学与应用生物学,2017,36(10):4380-4384.
[24] 唐媛,李春花,张瑗,等.蛋白质的二级结构预测研究进展[J].现代生物医学进展,2013,13(26):5180-5182.
[25] 赵伯阳,汪代华,徐刚毅,等.山羊CAST基因Ⅱ型转录本的克隆及在山羊不同组织中的表达[J].遗传,2011,33(4):358-364.
[26] 姚慧,陈智华,钟金城,等.CAPN1和CAST基因在牦牛不同组织中的表达差异研究[J].河南农业科学,2014,43(3):139-141,149.
[27] 刘安芳,朱庆,刘益平,等.钙蛋白酶抑制蛋白(CAST)基因在鸡不同组织和品种中的表达差异[J].中国农业科学,2007,40(10):2331-2335.

备注/Memo

备注/Memo:
收稿日期:2019-12-12 修回日期:2020-01-12
基金项目:山西省重点研发计划(国际科技合作)项目(201803D421022).
作者简介:李武峰(1967-),男,副教授.研究方向:动物遗传育种与繁殖.Email:leewf1967@163.com.通信作者杜敏(1969-),男,教授.研究方向:动物遗传育种与繁殖.Email:min.du@wsu.edu.
更新日期/Last Update: 2020-09-20