凡纳滨对虾TCP-1-eta基因的克隆及与耐寒性状的相关性

doi:10.3724/SP.J.1005.2011.00168

遗传 ›› 2011, Vol. 33 ›› Issue (2): 168-174.doi: 10.3724/SP.J.1005.2011.00168

凡纳滨对虾TCP-1-eta基因的克隆及与耐寒性状的相关性

殷勤1, 2, 彭金霞1, 崔亮1, 谢达祥1, 王志伟2, 李奎2, 陈晓汉1

1. 广西壮族自治区水产研究所, 南宁 530021;
2. 中国农业科学院北京畜牧兽医研究所, 北京 100193

收稿日期:2010-06-17 修回日期:2010-07-15 出版日期:2011-02-20 发布日期:2011-02-25
通讯作者: 陈晓汉 E-mail:chenxhan@yahoo.com.cn
基金资助:
国家科技支撑计划项目(编号：2008BADB9B03, 2007BAD29B01-6), 国家虾产业技术体系岗位科学家经费(编号：nycytx-46), 广西壮族自治区直属公益性科研院所基本科研业务费(编号：2060302 GXIF-2008-02, 2060302GXIF-2010-04)

Molecular cloning of Litopenaeus vannamei TCP-1-eta gene and analysis on its relationship with cold tolerance

YIN Qin^1,2, PENG Jin-Xia¹, CUI Liang¹, XIE Da-Xiang¹, WANG Zhi-Wei², LI Kui², CHEN Xiao-Han¹

1. Guangxi Fishery Institute, Nanning 530021, China;
2. Institute of Animal Sciences (IAS), Chinese Academy of Agricultural Sciences, Beijing 100193, China

Received:2010-06-17 Revised:2010-07-15 Online:2011-02-20 Published:2011-02-25
Contact: CHEN Xiao-Han E-mail:chenxhan@yahoo.com.cn

摘要/Abstract

摘要： 为研究凡纳滨对虾耐寒性状的分子机理, 文章克隆了凡纳滨对虾的耐寒相关基因TCP-1-eta并对其与耐寒性状的关系进行了研究。首先, 根据电子克隆所得序列设计引物, 采用RT-PCR方法克隆得到长1 705 bp的TCP-1-eta基因序列, 其中包括1 629 bp的完整开放阅读框, 编码542个氨基酸残基。然后, 运用荧光定量PCR对TCP-1-eta基因进行时空表达谱的分析: (1) 组织表达分析结果显示, 该基因在凡纳滨对虾肌肉组织中表达量最高; (2) 不同低温处理下的表达结果显示, 该基因在15℃开始呈上调表达, 13℃时表达量最高; (3) 13℃低温处理的表达结果显示, 该基因在36 h内呈小幅上调表达, 但36 h后表达量显著升高, 48 h表达量达到0 h的150倍。进一步采用PCR-RFLP方法对216只凡纳滨对虾TCP-1-eta基因进行了SNP多态性检测, 并将其与耐寒性状进行了关联分析。在该基因编码区第731碱基上发现C/T突变, 方差分析结果表明该位点的不同基因型与耐寒力指标CDH (Cooling-degree hours)值相关(P＜0.05), 其中CC基因型的耐寒能力比TT基因型强。

关键词: 凡纳滨对虾, TCP-1-eta, SNP, 耐寒性状

Abstract: To study the molecular mechanism of cold tolerance, we cloned TCP-1-eta homolog gene from Litopenaeus vannamei and studied its relationship with cold tolerance. Based on the sequence from electronic cloning, a pair of primers were designed, and a 1 705 bp cDNA was obtained by RT-PCR. The cDNA contains 1 629 bp ORF, which encodes a peptide of 542 aa. Tissue expression pattern by real-time PCR analysis revealed that TCP-1-eta mRNA mainly existed in muscle tissue. Cold induction by different temperatures revealed that TCP-1-eta mRNA expression started to increase after treatment at 15°C for 36 h, and the increase was remarkable after treatment at 13°C. As for treatment at 13°C for different times, the expression pattern was almost not changed during the first 36 h, but significantly increased after 36 h of treatment. A SNP (single nucleotide polymorphism) site was found in 731 bp of ORF, and the genotypes of 216 Litopenaeus vannamei were determined by PCR-RFLP. Variance analysis indicated that the SNP genotype was significantly related with cold tolerance parameter Cooling-Degree Hours (CDH), and CDH for CC type of Litopenaeus vannamei was longer than TT type.

Key words: Litopenaeus vannamei, TCP-1-eta gene, SNP, cold tolerance

殷勤，彭金霞，崔亮，谢达祥，王志伟，李奎，陈晓汉. 凡纳滨对虾TCP-1-eta基因的克隆及与耐寒性状的相关性[J]. 遗传, 2011, 33(2): 168-174.

YAN Qi, BANG Jin-Xia, CUI Liang, XIE Da-Xiang, WANG Zhi-Wei, LI Kui, CHEN Xiao-Han. Molecular cloning of Litopenaeus vannamei TCP-1-eta gene and analysis on its relationship with cold tolerance[J]. HEREDITAS, 2011, 33(2): 168-174.

参考文献

[1] Hightower LE. Heat shock, stress proteins, chaperones, and proteotoxicity. Cell, 1991, 66(2): 191-197.

[2] Hendrick JP, Hartl F. Molecular chaperone function of heat-shock proteins. Annu Rev Biohem, 1993, 62: 349-384.

[3] Welch WJ. How cells respond to stress. Sci Am, 1993, 268(5): 56-64.

[4] 程维杰, 李秋玲, 王长法, 王洪梅, 李建斌, 孙延鸣, 仲跻峰. 荷斯坦牛HSP70-1基因遗传多态性与乳腺炎抗性关系. 遗传, 2009, 31(2): 169-174.

[5] Sabahat S, Weiss D, Lurie S. The correlation between heat-shock protein accumulation and persistence and chill-ing tolerance in tomato fruit. Plant Physiol, 1996, 110(2): 531-537. [6] Sato A, Allona I, Collada C, Guevara MA, Casado R, Rodriguez-Cerezo E, Aragoncillo C, Gomez L. Heterologous expression of a plant small heat-shock protein enhances Esherichia coli viability under heat and cold stress. Plant Physiol, 1999, 120(2): 521-528.

[7] Kayukawa T, Chen B, Miyazake S, Shinoda T, Itoyama K, Ishikawa Y. Expression of mRNA for the t-complex polypeptide-1, a subunit of chaperonin CCT, is upregu-lated in association with increased cold hardiness in Delia antique. Cell Stress Chaperon, 2005, 10(3): 204-210.

[8] Campos ÉG, Hamdan FF. Cloning of the chaperonin t-complex polypeptide 1 gene from Schistosoma mansoni and studies of its expression levels under heat shock and oxidative stress. Parasitol Res, 2000, 86(3): 253-258.

[9] Chen ZZ, Cheng CC, Zhang JF, Cao LX, Chen L, Zhou LH, Jin YD, Ye Hua, Deng C, Dai ZH, Xu QH, Hu P, Sun SH, Shen Y, Chen LB. Transcriptomic and genomic evolution under constant cold in Antarctic notothenioid fish. Proc Natl Acad Sci USA, 2008, 105(35): 12944-12949.

[10] 张留所, 相建海.凡纳滨对虾微卫星位点在两个选育家系中遗传的初步研究. 遗传, 2005, 27(6) 919-924.

[11] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using realtime quantitative PCR and the 2-△△CT method. Methods, 2001, 25(4): 402-408.

[12] Chaani A, Gall GAE, Hulata G. Cold tolerance of tilapia species and hybrids. Aquacult Int, 2000, 8(4): 289-298.

[13] Yocum GD, Joplin KH, Denlinger DL. Expression of heat shock proteins in response to high and low temperature extremes in diapausing pharate larve of the gypsy moth, Lymanteia dispar. Arch Insect Biochem Physiol, 1991, 18(4): 239-249.

[14] Airaksinen S, Jokilehto T, Råbergh CMI, Nikinmaa M. Heat-and cold-inducible regulation of HSP70 expression in zebrafish ZF4 cells. Comp Biochem Physiol Part B, 2003, 136(2): 275-282.

[15] Huang LH, Wang CZ, Kang L. Cloning and expression of five heat shock protein genes in relation to cold hardening and development in the leafminer, Liriomyza sativa. J Insect Physiol, 2009, 55(3): 279-285.

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

凡纳滨对虾TCP-1-eta基因的克隆及与耐寒性状的相关性

Molecular cloning of Litopenaeus vannamei TCP-1-eta gene and analysis on its relationship with cold tolerance

PDF (PC)

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	郭晓媛, 孙昌春, 薛思瑶, 赵慧, 江丽, 李彩霞. 49AISNP：东亚北方三个族群遗传来源推断[J]. 遗传, 2021, 43(9): 880-889.
[2]	妥晓梅, 朱东丽, 陈晓峰, 荣誉, 郭燕, 杨铁林. 骨质疏松易感SNP rs4325274通过增强子远程调控SOX6基因的功能机制研究[J]. 遗传, 2020, 42(9): 889-897.
[3]	刘明, 李祎, 杨亚芳, 晏于文, 刘凡, 李彩霞, 曾发明, 赵雯婷. 中国汉族人群脸部特征相关SNP位点研究[J]. 遗传, 2020, 42(7): 680-690.
[4]	刘杨, 孙昌春, 马咪, 王玲, 赵雯婷, 马泉, 季安全, 刘京, 李彩霞. 74-plex SNPs复合检测体系在中国人群中的族群推断研究[J]. 遗传, 2020, 42(3): 296-308.
[5]	李智,何俊,蒋隽,Richard G. Tait Jr.,Stewart Bauck,过伟,吴晓林. 牛SNP芯片分型检出率和分型错误率对基因型填充准确率的影响[J]. 遗传, 2019, 41(7): 644-652.
[6]	何俊,钱长嵩,RichardG.TaitJr.,StewartBauck,吴晓林. SNP芯片数据估计动物个体基因组品种构成的方法及应用[J]. 遗传, 2018, 40(4): 305-314.
[7]	江丽,孙启凡,马泉,赵雯婷,刘京,赵蕾,季安全,李彩霞. 27-plex SNP种族推断方法的优化及验证[J]. 遗传, 2017, 39(2): 166-173.
[8]	刘娟, 孙艳, 姜强, 杨春红, 黄金明, 李建斌, 侯明海, 仲跻峰, 王长法, 刘保申. INCENP基因功能性单倍型可调控启动子活性并影响公牛精液品质[J]. 遗传, 2016, 38(1): 62-71.
[9]	钱旭丽, 曹新. 耳聋相关基因COCH的非同义单核苷酸多态性致聋表型预测[J]. 遗传, 2015, 37(7): 664-672.
[10]	李彩霞,贾竟,魏以梁,万立华,胡兰,叶健. 30个祖先信息位点的筛选及应用[J]. 遗传, 2014, 36(8): 779-785.
[11]	初芹, 李东, 侯诗宇, 石万海, 刘林, 王雅春. 基于DNA池测序法筛选奶牛高信息量SNP标记的可行性[J]. 遗传, 2014, 36(7): 691-696.
[12]	宋少娟, 郭亚平, 张学尧, 张建珍, 马恩波. 秀丽隐杆线虫抗铜突变体的筛选及SNP定位[J]. 遗传, 2014, 36(12): 1261-1268.
[13]	刘春,李琼艳,荀利杰,胡文波,吕金风,刘茹凤,夏庆友. 家蚕光泽小眼(varnished eye)突变基因的精细定位[J]. 遗传, 2014, 36(10): 1006-1012.
[14]	王洋坤, 胡艳, 张天真. RAD-seq技术在基因组研究中的现状及展望[J]. 遗传, 2014, 36(1): 41-49.
[15]	罗旭红刘志芳董长征. 基因水平的关联分析方法[J]. 遗传, 2013, 35(9): 1065-1071.