绵羊角蛋白关联蛋白8.2基因克隆与表达分析

doi:10.16431/j.cnki.1671-7236.2015.12.005

Abstract

Abstract: The object of this study was to clone the Keratin-associated protein 8.2 (KAP8.2) gene from Small-tail Han sheep and Xinji Fine-wool sheep and test the tissues expression pattern.In this study,the total RNA were extracted from different tissues firstly,and KAP8.2 gene was cloned by RT-PCR.In the end,the different expression patterns of KAP8.2 gene in Small-tail Han sheep and Xinji Fine-wool sheep was detected by QRT-PCR.The cloning results showed that the KAP8.2 gene were cloned,and the length of KAP8.2 mRNA was 574 bp,containing a 192 bp ORF which encoded a protein of 63 amino acids.These KAP8.2 gene belonged the typical HGT-KAP family because the rate of glycine and tyrosine were 23.8% and 20.6%,respectively.The polymorphism analysis showed that there were some polymorphisms in KAP8.2 gene and almost SNPs located on both sides of CDS region.The region which located on the 3'UTR c192+19-39 of KAP8.2 from Small-tail Han sheep might be the target of fru-let-7j.The expression pattern analysis showed that KAP8.2 gene was a multiple organs expression gene,and there were significant differences in expression patterns of KAP8.2 gene in Small-tail Han and Xinji Fine-wool sheeps.In Small-tail Han sheep,KAP8.2 gene was over-expressed in spleen,liver and skin,but in Xinji Fine-wool sheep,KAP8.2 gene was over-expressed in skin and heart.In conclusion,there were significant differences in gene polymorphisms and expression pattern between Small-tail Han sheep and Xinji Fine-wool sheep,and the result suggested the KAP8.2 gene associated with some phenotypes of wool.

Key words: KAP8.2 gene; Xinji Fine-wool sheep; Small-tail Han sheep; gene cloning

CLC Number:

Q786

ZHANG Li-chun, CAO Yang, SUN Fu-liang, YIN Feng, PIAO Qing-lin, WANG Xiao-yang, JIN Hai-guo, ZHANG Ming-xin. Cloning and Expression Pattern Analysis of KAP8.2 Gene in Sheep[J]. , 2015, 42(12): 3140-3145.

References

[1] Makar I A,Havryliak V V,Sedilo H M.Genetic and biochemical aspects of keratin synthesis by hair follicles[J].Tsitol Genet,2007,41(1):75-79.
[2] Rogers M A,Langbein L,Praetzel-Wunder S,et al.Human hair keratin-associated proteins (KAPs)[J].Int Rev Cytol,2006,251:209-263.
[3] Rogers G E,Powell B C.Organization and expression of hair follicle genes[J].J Invest Dermatol,1993,101(Suppl 1):50S-55S.
[4] Fratini A,Powell B C,Rogers G E.Sequence,expression,and evolutionary conservation of a gene encoding a glycine/tyrosine-rich keratin-associated protein of hair[J].J Biol Chem,1993,268(6):4511-4518.
[5] Marshall R C,Orwin D F,Gillespie J M.Structure and biochemistry of mammalian hard keratin[J].Electron Microsc Rev,1991,4(1):47-83.
[6] Li S W,Ouyang H S,Rogers G E,et al.Characterization of the structural and molecular defects in fibres and follicles of the Merino felting lustre mutant[J].Exp Dermatol,2009,18(2):134-142.
[7] Gong H,Zhou H,Hickford J G.Diversity of the glycine/tyrosine-rich keratin-associated protein 6 gene (KAP6) family in sheep[J].Mol Biol Rep,2011,38(1):31-35.
[8] Rogers M A,Langbein L,Winter H,et al.Characterization of a first domain of human high glycine-tyrosine and high sulfur keratin-associated protein (KAP) genes on chromosome 21q22.1[J].J Biol Chem,2002,277(50):48993-49002.
[9] Gong H,Zhou H,Dyer J M,et al.The sheep KAP8-2 gene,a new KAP8 family member that is absent in humans[J].Springerplus,2014,3:528.
[10] Schmittgen T D,Livak K J.Analyzing real-time PCR data by the comparative C(T) method[J].Nat Protoc,2008,3(6):1101-1108.
[11] 密冲,保灵燕,王莹,等.不同品种绵羊羊毛品质分析[J].畜牧与兽医,2012,44(12):30-31.
[12] 王蕾,张明新,王春昕,等.新吉细毛羊羊毛品质分析[J].中国草食动物科学,2012,S1:418-419.
[13] Huang C W,Li Y H,Hu S Y,et al.Differential expression patterns of growth-related microRNAs in the skeletal muscle of Nile tilapia (Oreochromis niloticus)[J].J Anim Sci,2012,90(12):4266-4279.
[14] Hertel J,Bartschat S,Wintsche A,et al.Evolution of the let-7 microRNA family[J].RNA Biol,2012,9(3):231-241.
[15] Yu Z,Gordon S W,Nixon A J,et al.Expression patterns of keratin intermediate filament and keratin associated protein genes in wool follicles[J].Differentiation,2009,77:307-316.
[16] Jin M,Wang L,Li S,et al.Characterization and expression analysis of KAP7.1,KAP8.2 gene in Liaoning new-breeding cashmere goat hair follicle[J].Mol Biol Rep,2011,38:3023-3028.
[17] 张桂山,薛景龙,孙丽敏,等.不同品种绵羊皮肤毛囊中角蛋白关联蛋白8-1基因差异表达研究[J].中国畜牧兽医,2014,41(5):76-79.

Cloning and Expression Pattern Analysis of KAP8.2 Gene in Sheep

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