Cloning and Temporal-spatial Expression Characteristics of GEM Gene in Tibetan Chicken

doi:10.7668/hbnxb.2018.02.007

Abstract

Abstract: The aim of this study was to clone GEM gene of Tibetan chicken,determine its expression profiling in different tissues and developmental stages,and analyzed the relationship between the expression level of GEM gene and intramuscular fat(IMF) deposition,which might provide basis for meat quality and fat deposition.1,81,119,154,210 days Tibetan chicken were selected. After slaughter,the tissue samples of heart,liver,spleen,lung,kidney,breast muscle,thigh muscle and subcutaneous fat were collected for the total RNA extraction. The Tibetan chicken GEM gene was cloned by Reverse Transcription PCR(RT-PCR),the quantitative Real-time quantitative PCR(qPCR) was used to detect the expression level of GEM gene in different tissues and developmental stages of breast muscle and thing muscle. The results showed that the full length of GEM gene was 940 bp(GenBank accession number:KY747399),containing 894 bp CDS,24 bp 5'UTR and 22 bp 3'UTR,encoding 297 amino acids. The Tibetan chicken GEM protein shared higher similarity(83.50%-98.32%) with Gallus gallus, Anas platyrhynchos, Homo sapiens and Mus musculu. The tissues expression analysis showed that GEM gene kept the highest expression level in lung of Tibetan chicken,which was remarkably higher than other tissues(P<0.01),and also higher in spleen and subcutaneous fat. The temporal expression showed that the expression pattern of GEM gene was different in breast muscle and thigh muscle of Tibetan chicken in different stages. The expression level of GEM mRNA in breast muscle decreased with the increasing daily age,however,in thigh muscle increased first and then decreased with Tibetan chicken growth,and peaked at the daily age of 119. The expression levels of Tibetan chicken GEM gene in breast muscle and thigh muscle at different developmental stages presented unlike relationship with IMF content,and there were genders differences. And the correlation in thigh muscle of male Tibetan chicken was remarkably stronger(r=0.414, P<0.05). These data indicate that GEM gene may play an important role in lipid metabolism of Tibetan chicken.

Key words: Tibetan chicken, GEM gene, Tissue expression, Temporal expression, Intramuscular fat content

CLC Number:

Q78
S831.03

XU Qing, LIN Sen, XU Yaou, ZHU Jiangjiang, WANG Yong, LIN Yaqiu. Cloning and Temporal-spatial Expression Characteristics of GEM Gene in Tibetan Chicken[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2018, 33(2): 42-48. doi: 10.7668/hbnxb.2018.02.007.

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References

[1] 聂晓庆,林亚秋,徐亚欧,等.藏鸡磷酸酪氨酸互作结构域1(PID1)的克隆及组织表达谱研究[J].畜牧兽医学报,2016,47(6):1102-1111.
[2] 王翔宇.藏鸡胚胎低氧适应的转录组差异表达分析[D].北京:中国农业大学,2014.
[3] 林亚秋,徐亚欧,张润锋,等.藏鸡Chemerin和ChemeR23基因时序表达及其与肌内脂肪含量的相关性研究[J].畜牧兽医学报,2015,46(8):1290-1299.
[4] 甘麦邻,堵晶晶,杨琼,等.动物肌内脂肪对肉质的影响及其分子机制研究进展[J].现代畜牧兽医,2017(10):51-57.
[5] Piddini E,Schmid J A,De Martin R,et al.The Ras-like GTPase Gem is involved in cell shape remodelling and interacts with the novel kinesin-like protein KIF9[J].The EMBO Journal,2001,20(15):4076-4087.
[6] Maguire J,Santoro T,Jensen P,et al.Gem:an induced,immediate early protein belonging to the Ras family[J].Science,1994,265(5169):241-244.
[7] Opatowsky Y,Sasson Y,Shaked I,et al.Structure-function studies of the G-domain from human gem,a novel small G-protein[J].FEBS Letters,2006,580(25):5959-5964.
[8] Andrieu G,Quaranta M,Leprince C,et al.Gem GTPase acts upstream Gmip/RhoA to regulate cortical actin remodeling and spindle positioning during early mitosis[J].Carcinogenesis,2014,35(11):2503-2511.
[9] Ward Y,Yap S F,Ravichandran V,et al.The GTP binding proteins Gem and Rad are negative regulators of the Rho-Rho kinase pathway[J].Journal of Cell Biology,2002,157(2):291-302.
[10] 龚海霞,郭锡熔,陈荣华,等.肥胖大鼠脂肪组织基因表达谱系研究[J].中华医学遗传学杂志,2003,20(3):268-271.
[11] 刘勇,郭锡熔,潘晓勤,等.GEM基因在3T3-L1脂肪细胞诱导分化过程中的表达变化[J].现代诊断与治疗,2005,16(5):265-268.
[12] Li Q,Zuo L L,Lin Y Q,et al.Cloning and expression of SFRP5 in tibetan chicken and its relationship with IMF deposition[J].Animal Biotechnology,2016,27(4):231-237.
[13] 林森,林亚秋,朱江江,等.藏鸡FTO基因表达的时空特性及与肌内脂肪含量的相关分析[J].畜牧兽医学报,2017,48(7):1191-1201.
[14] Ward Y,Spinelli B,Quon M J,et al.Phosphorylation of critical serine residues in gem separates cytoskeletal reorganization from down-regulation of Calcium Channel activity[J].Molecular and Cellular Biology,2004,24(2):651-661.
[15] Hatzoglou A,Ader I,Splingard A A,et al.Gem associates with ezrin and acts via the Rho-GAP protein gmip to down-regulate the Rho pathway[J].Molecular Biology of the Cell,2007,18(4):1242-1252.
[16] Ward Y,Kelly K.Gem protein signaling and regulation[J].Methods Enzymol,2006,407:468-483.
[17] Gunton J E,Sisavanh M,Stokes R A,et al.Mice deficient in GEM GTPase show abnormal glucose homeostasis due to defects in Beta-Cell Calcium handling[J].PLoS One,2012,7(6):e39462.
[18] 石佰丽,吕梦燕,赵秋雁.小GTPase蛋白家族的研究进展[J].安徽农业科学,2012,40(5):2566-2569,2573.
[19] Kimj H,Lee J,Oh B,et al.Prediction of phosphorylation sites using SVMs[J].Bioinformatics,2004,20(17):3179-3184.
[20] Gupta R,Brunak S.Prediction of glycosylation across the human proteome and the correlation to protein function[J].Pac Symp Biocomput,2002,7:310-322.

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