Please wait a minute...
浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2019, Vol. 45 Issue (2): 237-242    DOI: 10.3785/j.issn.1008-9209.2018.03.131
动物科学与动物医学     
鸡GNAS基因c.36-2277T＞C突变对萤光素酶表达的影响
王欢欢(),张雷,葛莹,宋丹丹,楼立峰,章学东()
杭州市农业科学研究院，杭州 310024
Effect of c.362277T C mutation on the expression of luciferase in chicken GNAS gene
Huanhuan WANG(),Lei ZHANG,Ying GE,Dandan SONG,Lifeng LOU,Xuedong ZHANG()
Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China
 全文: PDF(3061 KB)   HTML ( )  HTML
摘要：

为分析探讨鸡GNAS基因ENSGALT00000062075.1:c.36-2277T＞C突变对转录调控的影响，本试验选择乌骨鸡和芦花鸡血液样本，构建了c.36-2277T＞C突变点的不同基因型质粒，并转染鸡胚成纤维细胞后，检测双萤光素酶报告基因分析系统的表达。结果表明：样本芦花鸡均为TT基因型，乌骨鸡均为CC基因型；c.36-2277T＞C突变点的上下游－175～+818 bp片段具有显著增强转录活性作用（P＜0.01），且CC基因型活性极显著高于TT基因型（P＜0.000 1）。转录因子预测结果表明：此993 bp片段中TGGCA结合蛋白位点数量最多；紧邻突变点分别为AP-1和ER-alpha/T3R-alpha结合位点。研究认为，GNAS基因c.36-2277T＞C突变点的碱基类型会对基因转录调控进程产生重要影响，且该突变可能是通过与上下游转录因子结合位点共同作用来发挥效用。
关键词： 鸡;  GNAS基因;  突变;  萤光素酶    
Abstract:

In order to analyze the effect of ENSGALT00000062075.1:c.36-2277T＞C mutation on the expression of transcriptional regulation in chicken GNAS gene, this study collected blood DNA samples from black-bone chicken and barred chicken to construct different genotypic plasmids of c.36-2277T＞C mutation, and detected the expression of dual-luciferase reporter analysis system after transfecting chicken embryo fibroblasts. The result showed that all barred chicken samples belonged to TT genotypes and all black-bone chicken samples were CC genotypes. The section from 175 bp upstream to 818 bp downstream of c.36-2277T＞C mutation had significantly transcript enhancing activity (P＜0.01). The activity of CC genotype was significantly higher than that of the TT genotype (P＜0.000 1). The result of transcript factor prediction showed that the number of TGGCA-binding protein sites was the largest in this 993 bp section, and the adjacent mutations were AP-1 and ER-alpha/T3R-alpha binding sites. The study suggests that the base types of c.36-2277T＞C mutation in GNAS gene have an important impact on the process of gene transcription regulation, and this mutation plays the role associated with upstream and downstream binding sites of transcript factors.
Key words: chicken    GNAS gene    mutation    luciferase
收稿日期: 2018-03-13 出版日期: 2019-04-25
CLC:  S 831.2  
基金资助: 杭州市科学技术委员会农业科研主动设计项目(20162012A04)
通讯作者: 章学东     E-mail: wennuan2009@163.com;bigzhengliang@hotmail.com
作者简介: 王欢欢（https://orcid.org/0000-0002-8778-0841），Tel: +86-571-87093912，E-mail： wennuan2009@163.com|章学东（https://orcid.org/0000-0002-1347-3136），Tel: +86-571-87310564，E-mail： bigzhengliang@hotmail.com
服务　　
把本文推荐给朋友
加入引用管理器
E-mail Alert
RSS
作者相关文章　　
王欢欢
张雷
葛莹
宋丹丹
楼立峰
章学东

引用本文:

王欢欢,张雷,葛莹,宋丹丹,楼立峰,章学东. 鸡GNAS基因c.36-2277T＞C突变对萤光素酶表达的影响[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(2): 237-242.

Huanhuan WANG,Lei ZHANG,Ying GE,Dandan SONG,Lifeng LOU,Xuedong ZHANG. Effect of c.362277T C mutation on the expression of luciferase in chicken GNAS gene. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(2): 237-242.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2018.03.131    或    http://www.zjujournals.com/agr/CN/Y2019/V45/I2/237

图1  基因组突变位置及测序结果
图2  质粒PCR电泳结果
图3  萤光素酶活性检测结果
图4  突变点上下游993 bp碱基序列及转录因子
1 WEINSTEIN L S , LIU J , SAKAMOTO A , et al . Minireview: GNAS: normal and abnormal functions. Endocrinology, 2004,145(12):5459-5464.
2 BOYCE A M , FLORENZANO P , CASTRO L F DE , et al . Fibrous dysplasia/McCune-Albright syndrome. Seattle, US: University of Washington, 2015:1993-2019.
3 LEMOS M C , THAKKER R V . GNAS mutations in pseudohypoparathyroidism type 1a and related disorders. Human Mutation, 2015,36(1):11-19.
4 王欢欢,陈美玲,楼立峰,等 .鸡GNAS基因启动子突变及其与肤色性状的相关性.畜牧兽医学报,2016,47(12):2354-2361.
WANG H H , CHEN M L , LOU L F , et al . GNAS gene promoter mutation in chicken and the correlation with skin color traits. Acta Veterinaria et Zootechnica Sinica, 2016,47(12):2354-2361. (in Chinese with English abstract)
5 OGINO S , GULLEY M L , DUNNEN J T DEN , et al . Standard mutation nomenclature in molecular diagnostics. Journal of Molecular Diagnostics, 2007,9(1):1-6.
6 WEINSTEIN L S , SHENKER A , GEJMAN P V , et al . Activating mutations of the stimulatory G protein in the McCune-Albright syndrome. The New England Journal of Medicine, 1991,325:1688-1695.
7 COLLINS M T , SARLIS N J , MERINO M J , et al . Thyroid carcinoma in the McCune-Albright syndrome: contributory role of activating Gs α mutations. The Journal of Clinical Endocrinology & Metabolism, 2003,88(9):4413-4417.
8 NGUYEN L P , PAN J L , DINH T T , et al . Role and species-specific expression of colon T cell homing receptor GPR15 in colitis. Nature Immunology, 2014,16:207-213.
9 SOLDNER F , STELZER Y , SHIVALILA C S , et al . Parkinson-associated risk variant in distal enhancer of α-synuclein modulates target gene expression. Nature, 2016,533:95-99.
10 MURTAGH J , MARTIN F , GRONOSTAJSKI R M . The nuclear factor I (NFI) gene family in mammary gland development and function. Journal of Mammary Gland Biology and Neoplasia, 2003,8(2):241-254.
11 TIAN T V , GRAF T . CEBPA (CCAAT/enhancer binding protein (C/EBP), alpha). Atlas of Genetics and Cytogenetics in Oncology and Haematology, 2015,19(4):249-255.
12 COCKERILL P N , BERT A G , JENKINS F , et al . Human granulocyte-macrophage colony-stimulating factor enhancer function is associated with cooperative interactions between AP-1 and NFATp/c. Molecular and Cellular Biology, 1995,15(4):2071-2079.
13 YEH J H, LECINE P , NUNES J A , et al . Novel CD28-responsive enhancer activated by CREB/ATF and AP-1 families in the human interleukin-2 receptor α-chain locus. Molecular and Cellular Biology, 2001,21(14):4515-4527.
[1] 侯福银,杨智青,陈长宽,时凯,金崇富,陈应江. 原花青素对肉鸡肠道及屠宰性能、脂质代谢和肉品质的影响[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(1): 119-125.
[2] 张丽, 刘丽霞, 戴洪伟, 陈红, 王瑞, 岳炳辉. 静宁鸡肌肉生长抑制素基因单核苷酸多态性筛查及生物信息学分析[J]. 浙江大学学报（农业与生命科学版）, 2018, 44(5): 629-637.
[3] 张志昌, 潘伟槐, 严旭, 尹守鹏, 程祝宽, 潘建伟. 水稻根系生长素抗性突变体的筛选及表型鉴定[J]. 浙江大学学报（农业与生命科学版）, 2018, 44(3): 311-317.
[4] 余道兵, 程学松, 王群, 史艳可, 张昕. 植酸酶基因的定点突变及其在巴斯德毕赤酵母表面展示[J]. 浙江大学学报（农业与生命科学版）, 2018, 44(1): 10-20.
[5] 徐红蕊, 陈小连, 时建青, 宋琼莉. 艾叶对热应激蛋鸡抗氧化功能、产蛋性能和蛋品质的影响[J]. 浙江大学学报(农业与生命科学版), 2017, 43(1): 113-119.
[6] 陆俊贤, 贾晓旭, 唐修君, 樊艳凤, 唐梦君, 高玉时, 苏一军. 2个云南原始鸡种遗传多样性及其与红色原鸡的亲缘关系[J]. 浙江大学学报(农业与生命科学版), 2016, 42(3): 385-390.
[7] 陈婷婷，李显富，贾春雷，安雪琴，朱俊，肖立增，戴秀梅，张建奎. 云烟87突变体新品系的特征特性及其DNA指纹图谱鉴定[J]. 浙江大学学报(农业与生命科学版), 2016, 42(2): 179-189.
[8] 陈林，杨新国，宋乃平，翟德苹，赵伟，刘学东. 中间锦鸡儿根水浸提液对7种灌草的化感作用及其化学成分分析[J]. 浙江大学学报(农业与生命科学版), 2016, 42(2): 150-162.
[9] 朱开元, 刘慧春, 周江华, 邹清成, 马广莹, 张加强, 魏红旭. 延长光周期对罗汉松和鸡爪槭苗期生长及养分吸收利用的影响[J]. 浙江大学学报(农业与生命科学版), 2016, 42(2): 190-198.
[10] 刘媛, 林曼曼, 张霄, 徐重新, 焦凌霞, 仲建锋, 武爱华, 刘贤金. 基因突变技术在抗体亲和力体外成熟中的应用[J]. 浙江大学学报(农业与生命科学版), 2016, 42(1): 1-7.
[11] 郑嫩珠, 李丽, 辛清武, 缪中纬, 朱志明, 刘凤辉, 吴俭飞, 卢立志. 内参基因对TYR、MITF和ASIP基因在白绒乌骨鸡各组织表达水平的影响[J]. 浙江大学学报(农业与生命科学版), 2015, 41(6): 732-740.
[12] 童杰鹏,童川,王艳,任三娟,沈圣泉. 水稻低纤维突变体LCM527-1性状鉴定、遗传分析与基因定位[J]. 浙江大学学报(农业与生命科学版), 2015, 41(03): 261-268.
[13] 翟德苹, 陈林, 杨明秀, 宋乃平, 杨新国. 荒漠草原不同生长年限中间锦鸡儿灌丛枝系构型特征[J]. 浙江大学学报(农业与生命科学版), 2015, 41(03): 340-348.
[14] 袁兵， 崔海瑞， 富昊伟， 蒋萌， 李瑞清， 赵海军， 舒庆尧. 水稻Os09g24220基因插入突变体的分子鉴定与农艺性状分析[J]. 浙江大学学报(农业与生命科学版), 2014, 40(4): 456-462.
[15] 葛延松， 曹嫦妤， 王丽丽， 李楠， 江秀清， 李金龙*. 鸡硒蛋白T的硒代半胱氨酸插入序列元件、蛋白结构与功能及组织表达差异[J]. 浙江大学学报(农业与生命科学版), 2014, 40(1): 9-15.