Mengtig ZHAI, Jian MOU, Mengtig ZHU, Yanping LIANG, Mingyuan WANG, ? Zongsheng 1 ZHAO, Hongmei ZHANG

Polymorphisms of MBL Gene Introns and Their Association with MBL Serum Levels in Hu Sheep

PCR single-strand conformation polymorphism (SSCP) and DNA sequencing techniques were used to analyze the genetic polymorphism of mannose-binding lectin (MBL) gene 3 introns in Hu sheep. The results showed that 3 introns of MBL gene had polymorphism, 3 genotypes were identified in intron1, which were controlled by 2 alleles, respectively; 3 genotypes were identified in intron2, which were controlled by 2 alleles, respectively; 3 genotypes were identified in intron3, which were controlled by 2 alleles, respectively; 6 new single nucleotide polymorphisms(SNPs) mutation sites were found, g.288T>A in intron1, g. 1091 T>C, g.1096A>C, g.1770G>C in intron 2, g.2297C>T, g.2331G>A in intron3. Use ELISA to detect MBL serum level in Hu sheep, then use One-way ANOVA analyze the relationship of diffrent genotypes and MBL serum level. And the result showed that the MBL serum level of AA genotype was higher than BB genotype (P

Hu Koyunlarında MBL Gen İntronlarının Polimorfizmi ve MBL Serum Seviyeleri ile İlişkisi

PCR single-strand conformation polymorphism (SSCP) ve DNA sekanslama teknikleri, Hu koyunlarında mannoz bağlayan lektin (MBL) gen 3 intronunda genetik polimorfizmi analiz etmede kullanılmıştır. Sonuçlar MBL geninin 3 intronunda polimorfizmin olduğunu gösterdi, ve intron 1’de sırayla 2 allel tarafından kontrol edilen 3 genotip, intron 2’de sırayla 2 allel tarafından kontrol edilen 3 genotip, intron 3’de de sırayla 2 allel tarafından kontrol edilen 3 genotip tespit edildi. 6 yeni tek nükleotid polimorfizm (SNPs) mutasyon alanı (g.288T>A intron 1’de, g. 1091 T>C, g.1096A>C, g.1770G>C intron 2’de, g.2297C>T, g.2331G>A intron 3’de) bulundu. Hu koyunlarında MBL serum seviyelerini belirlemek amacıyla ELİSA uygulandı, sonrasında Tek yönlü ANOVA kullanılarak farklı genotipler ile MBL serum seviyeleri arasındaki ilişki analiz edildi. Sonuçlar intron 1’de AA genotipinin MBL serum seviyesinin BB genotipinden daha fazla olduğunu (P

PDF

___

Degn SE, Thiel S: Humoral pattern recognition and the complement system. Scand J Immunol, 78 (2): 181-193, 2013. DOI: 10.1111/sji.12070

Wang M, Zhang Y, Chen Y, Zhang L, Lu X, Chen Z: Mannan-binding lectin regulates dendritic cell maturation and cytokine production induced by lipopolysaccharide. BMC Immunol, 12:1, 2011. DOI: 10.1186/1471- 2172-12-1

Zhao F, Zhao Z, Yan G, Wang D, Ban Q, Yu P, Zhang W, Luo Y: Polymorphisms in mannose-binding lectin (MBL) gene and their association with MBL protein levels in serum in the Hu sheep. Vet Immunol Immunopathol, 140, 297-302, 2011. DOI: 10.1016/j.vetimm.2010.12.009

Smithson A, Perello R, Aibar J, Espinosa G, Tassies D, Freire C, Castro P, Suarez B, Lozano F, Nicolas JM: Genotypes coding for low serum levels of mannose-binding lectin are underrepresented among individuals suffring from noninfectious systemic inflmmatory response syndrome. Clin Vaccine Immunol, 17 (3): 447-453, 2010. DOI: 10.1128/ CVI.00375-09

Turner MW: The role of mannose binding lectin in health and disease. Mol Immunol, 40, 423-429, 2003. DOI: 10.1016/S0161-5890(03)00155-X

Madsen HO, Garred P, Kurtzhals JAL, Lamm LU, Ryder LP, Thiel S, Svejgaard A: A new frequent allele is the missing link in the structural polymorphism of the human mannan-binding protein. Immunogenetics. 40, 37-44, 1994. DOI: 10.1007/BF00163962

Sumiya MTabona P, Arai T, Summerfield JA, Super M, Levinsky RJ, Turner MW: Molecular basis of opsonic defect in immunodeficient children. Lancet, 337, 1569-1570, 1991. DOI: 10.1016/0140-6736(91)93263-9

Lipscombe RJ, Sumiya M, Summerfield JA, Turner MW: Distinct physicochemical characteristics of human mannose binding protein expressed by individuals of diffring genotype. Immunology, 85, 660-667, 1995.

Wang HM, Zhang LB, Hou MH, Wang CF, Wang LL, Sun T, He HB, Zhong JF: Cloning and activity analysis of bovine natural resistance associated macrophage protein1(Nramp 1)gene promoter. Scicntia Agric Sinica, 21, 2011.

Ghang X, Luo J, Li JH, et al: Cloning and activity dct-urination of fatty acid synthasc gene promoter of xinong saancn dairy goat. Scicntia Agric Sinica, 28, 2010.

Sambrook J, Russell DW: Molecular Cloning: A Laboratory Manual. Vol. 3, 3 rd ed., Cold Spring Harbor Laboratory Press, New York, 49-56, 2001.

Lan XY, Pan CY, Chen H, Zhao M, Li JY, Yu J, Zhang CL, Lei CZ, Hua LS, Yang XB: The novel SNPs of the IGFBP3 gene and their associations with litter size and weight traits in goat. Arc Anim Breed, 50, 223-224,2007.

Gadjeva M, Takahashi K, Thiel S: Mannan-binding lectin-a soluble pattern recognition molecule. Mol Immunol, 41 (2-3): 113-121, 2004. DOI: 10.1016/j.molimm.2004.03.015

Thiel S, Frederiksen PD, Jensenius JC: Clinical manifestations of mannan-binding lecain deficiency. Mol Immunol, 43 (1-2): 86-96, 2006. DOI: 10.1016/j.molimm.2005.06.018

Chatterjee A, Rathore A, Yamamoto N, Dhole TN: Mannose-binding lectin (+54) exon-1 gene polymorphism influence human immunodeficiency virus-1 susceptibility in North Indians. Tissue Antigens, 77, 18- 22, 2011. DOI: 10.1111/j.1399-0039.2010.01563.x

Michelow IC, Lear C, Scully C, Prugar LI, Longley CB, Yantosca LM, Ji X, Karpel M, Brudner M, Takahashi K, Spear GT, Ezekowitz RA, Schmidt EV, Olinger GG: High-dose mannose-binding lectin therapy for Ebola virus infection. J Infect Dis, 203, 175-179, 2011. DOI: 10.1093/infdis/ jiq025

Storgaard P, Nielsen EH, Andersen O, Skriver E, Mortensen H, Hojrup P, Leslie G, Holmskow U, Svehag SE: Isolation and characterization of porcine mannan-binding proteins of diffrent size and ultrastructure. Scand J Immunol, 43, 289-296, 1996. DOI: 10.1046/ j.1 365-3083.1996.d01-39.x