延边黄牛转铁蛋白受体2基因克隆与序列分析

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doi:10.3864/j.issn.0578-1752.2012.10.015

摘要： 【目的】克隆延边黄牛转铁蛋白受体2（transferrin receptor 2，TFR2）基因，并分析该基因在不同组织器官中的表达分布。【方法】通过提取肝脏组织总RNA，采用RT-PCR及RACE方法克隆延边黄牛TfR2基因，采用生物信息学软件进行序列分析，用半定量PCR（SqRT-PCR）方法分析TfR2基因在不同组织器官中的表达分布规律。【结果】①成功克隆出了延边黄牛TfR2基因完整ORF区及3′UTR区（GenBank登录号：GU553087），该基因全长2 901 bp，ORF区2 412 bp，编码803个氨基酸；②延边黄牛TfR2基因核苷酸序列与其它物种的同源性在80%以上，不同物种间TfR2基因氨基酸序列，尤其是影响其功能的关键氨基酸位点高度保守；③各物种TfR2蛋白功能结构域同样高度保守；④牛TfR2基因主要在肝脏中表达，其它组织，如脾脏、心脏、肾脏和肠道（十二指肠）中也存在少量表达。【结论】不同物种间TfR2基因具有高度同源性，功能结构域及组织分布规律具有高度保守性。

关键词: 延边黄牛, 转铁蛋白受体2基因, RACE

Abstract: 【Objective】 The transferrin receptor 2 (TfR2) gene was cloned from Yanbian Yellow cattle, the sequence was analyzed by bioinformatics software and the tissue expression distribution was tested by semi-quantitative reverse transcription and polymerase chain reaction (SqRT-PCR).【Method】 The total RNA was extracted from Yanbian Yellow cattle liver. The TfR2 gene was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The TfR2 gene was analyzed by bioinformatic softwares and the TfR2 gene tissue distribution was tested by SqRT-PCR in the end. 【Result】 The complete CDs and 3′ UTR region of TfR2 gene was cloned (GenBank accession: GU553087). The TfR2 gene was 2901 bp in length and contained a 2412 -nucleotides-long open reading frame (ORF) which encoded a protein of 803 amino acid residues. The sequence analysis result showed that Yanbian Yellow cattle TfR2 gene has over 80% homology with mammals in nucleotides, the amino acid sequences of TfR2 gene from different mammals were conserved, especially some amino acid sites which affect its function in human were highly conserved. The protein functional domain analysis showed that TfR2 gene from different mammals were highly conserved. The expression profiles analysis showed that the TfR2 gene was highly expressed in liver, constitutively expressed in spleen, heart, kidney and intestine (duodenum).【conclusion】 The TfR2 gene from Yanbian Yellow cattle is highly homologous with other mammals, the functional domain and tissue distribution are highly conserved in different mammals.

Key words: Yanbian Yellow cattle, TfR2 gene, RACE

张立春, 金海国, 李赵志, 任春宇, 曹阳, 周国利, 金鑫. 延边黄牛转铁蛋白受体2基因克隆与序列分析[J]. 中国农业科学, 2012, 45(10): 2022-2030.

ZHANG Li-Chun, JIN Hai-Guo, LI Zhao-Zhi, REN Chun-Yu, CAO Yang, ZHOU Guo-Li, JIN Xin. Cloning and Sequence Analysis of the Transferrin Receptor 2 Gene from Yanbian Yellow Cattle[J]. Scientia Agricultura Sinica, 2012, 45(10): 2022-2030.

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[1]Theil E C. Iron, ferritin, and nutrition. Annual Review Nutrition, 2004, 24: 327-343.

[2]Valerio L G. Mammalian iron metabolism. Toxicology Mechanisms and Methods, 2007, 17(9): 497-517.

[3]Conrad M E, Umbreit J N. Pathways of iron absorption. Blood Cells, Molecules, and Diseases, 2002, 29(3): 336-355.

[4]Hemadi M, Kahn P H, Miquel G, El Hage Chahine J M. Transferrin′s mechanism of interaction with receptor 1. Biochemistry, 2004, 43(6): 1736-1745.

[5]Kawabata H, Yang R, Hirama T, Vuong P T, Kawano S, Gombart A F, Koeffler H P. Molecular cloning of transferrin receptor 2. A new member of the transferrin receptor-like family. Journal of Biology Chemistry, 1999, 274(30): 20826-20832.

[6]Kawabata H, Germain R S, Ikezoe T, Tong X, Green E M, Gombart A F, Koeffler H P. Regulation of expression of murine transferrin receptor 2. Blood, 2001, 98(6): 1949-1954.

[7]Hentze M W, Muckenthaler M U, Andrews N C. Balancing acts: molecular control of mammalian iron metabolism. Cell, 2004, 117(3): 285-297.

[8]Chiotoglou I, Smilevska T, Samara M, Likousi S, Belessi C, Athanasiadou I, Stavroyianni N, Samara S, Laoutaris N, Vamvakopoulos N, Anagnostopoulos A, Fassas A, Stamatopoulos K, Kollia P. Predominantly post-transcriptional regulation of activation molecules in chronic lymphocytic leukemia: the case of transferrin receptors. Blood Cells, Molecules, and Diseases, 41(2): 203-209.

[9]Mattman A, Huntsman D, Lockitch G, Langlois S, Buskard N, Ralston D, Butterfield Y, Rodrigues P, Jones S, Porto G, Marra M, de Sousa M, Vatcher G. Transferrin receptor 2 (TfR2) and HFE mutational analysis in non-C282Y iron overload: identification of a novel TfR2 mutation. Blood, 2002, 100(3): 1075-1077.

[10]Roetto A, Daraio F, Alberti F, Porporato P, Cali A, de Gobbi M, Camaschella C. Hemochromatosis due to mutations in transferrin receptor 2. Blood Cells, Molecules, and Diseases, 2002, 29(3): 465-470.

[11]Wilson M D, Riemer C, Martindale D W, Schnupf P, Boright A P, Cheung T L, Hardy D M, Schwartz S, Scherer S W, Tsui L C, Miller W, Koop B F. Comparative analysis of the gene-dense ACHE/TFR2 region on human chromosome 7q22 with the orthologous region on mouse chromosome 5. Nucleic Acids Research, 2001, 29(6): 1352-1365.

[12]Fleming R E, Ahmann J R, Migas M C, Waheed A, Koeffler H P, Kawabata H, Britton R S, Bacon B R, Sly W S. Targeted mutagenesis of the murine transferrin receptor-2 gene produces hemochromatosis. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(16): 10653-10658.

[13]Hattori A, Wakusawa S, Hayashi H, Harashima A, Sanae F, Kawanaka M, YamadaG, Yano M, Yoshioka K. AVAQ 594-597 deletion of the TfR2 gene in a Japanese family with hemochromatosis. Hepatology Research, 2003, 26(2): 154-156.

[14]Chen J, Chloupkova M. Abnormal iron uptake and liver cancer. Cancer Biology and Therapy, 2009, 8(18): 1699-1708.

[15]Calzolari A, Oliviero I, Deaglio S, Mariani G, Biffoni M, Sposi N M, Malavasi F, Peschle C, Testa U. Transferrin receptor 2 is frequently expressed in human cancer cell lines. Blood Cells, Molecules, and Diseases, 2007, 39(1): 82-91.

[16]林亚秋, 郑玉才, 吉红, 贺庆华, 邱翔. 草鱼转铁蛋白受体cDNA序列克隆及其组织表达差异. 湖北农业科学, 2009, 48(6): 1289-1292.

Lin Y Q, Zheng Y C, Ji H, He Q H, Qiu X. Cloning of TfR partial cDNA sequence in grass carp and its tissue differential expression. Huibei Agricultural Sciences, 2009, 48(6): 1289-1292. (in Chinese)

[17]高明英, 朱新平, 赵密, 史燕, 赵建. 黄喉拟水龟转铁蛋白基因的克隆以及表达特征分析. 水生生物学报, 2011, 35(4): 557-564.

Gao M Y, Zhu X P, Zhao M, Shi Y, Zhao J. Cloning and expression pattern analysis of transferrin gene in yellow pond turtle. Acta Hydrobiologica Sinica, 2011, 35(4): 557-564. (in Chinese)

[18]Camaschella C. Why do humans need two types of transferrin receptor? Lessons from a rare genetic disorder. Haematologica, 2005, 90(3): 296-928.

[19]Kumar S, Nei M, Dudley J, Tamura K. MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Briefings in Bioinformatics, 2008, 9(4): 299-306.

[20]Lambert L A, Mitchell S L. Molecular evolution of the transferrin receptor/glutamate carboxypeptidase Ⅱ family. Journal of Molecular Evolution, 2007, 64(1): 113-128.

[21]鞠志花, 李秋玲, 黄金明, 王霁, 李荣岭, 李建斌, 仲跻峰, 王长法. 中国荷斯坦牛转铁蛋白基因SNPs的检测及其与产奶性能的关系. 中国农业科学, 2011, 44(14): 3027-3035.

Ju Z H, Li Q L, Huang J M, Wang J, Li R L, Li J B, Zhong J F, Wang C F. SNPs detection in Chinese holstein cattle transferrin gene and its associations with milk performance. Scientia Agricultura Sinica, 2011, 44(14): 3027-3035. (in Chinese)

[22]张永宏, 潘英树, 高妍, 马倩, 刘同欣, 秦莹, 赵志辉, 张嘉保. 草原红牛转铁蛋白和后转铁蛋白多态性及其与生产性能相关性研究. 农业科学与技术, 2008, 9(5): 109-112.

Zhang Y H, Pan Y S, Gao Y, Ma Q, Liu T X, Qin Y, Zhao Z H, Zhang J B. Studies on transferrin and posttremsferr polymorphism and their relationship with performances in red steppe. Agricultural Science and Technology, 2008, 9(5): 109-112. (in Chinese)

[23]Richardson D R, Ponka P. The molecular mechanisms of the metabolism and transport of iron in normal and neoplastic cells. Biochimica et Biophysica Acta, 1997, 1331(1): 1-40.

[24]Fleming R E, Migas M C, Holden C C, Waheed A, Britton R S, Tomatsu S, Bacon B R, Sly W S. Transferrin receptor 2: continued expression in mouse liver in the face of iron overload and in hereditary hemochromatosis. Proceedings of the National Academy of Sciences of the United States of America, 2000, 97(5): 2214-2219.

[25]Deaglio S, Capobianco A, Cali A, Bellora F, Alberti F, Righi L, Sapino A, Camaschella C, Malavasi F. Structural, functional, and tissue distribution analysis of human transferrin receptor-2 by murine monoclonal antibodies and a polyclonal antiserum. Blood, 2002, 100(10): 3782-3789.

[26]Feder J N, Penny D M, Irrinki A, Lee V K, Lebron J A, Watson N, Tsuchihashi Z, Sigal E, Bjorkman P J, Schatzman R C. The hemochromatosis gene product complexes with the transferrin receptor and lowers its affinity for ligand binding. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95(4): 1472-1477.

[27]Camaschella C, Roetto A, Cali A, de Gobbi M, Garozzo G, Carella M, Majorano N, Totaro A, Gasparini P. The gene TFR2 is mutated in a new type of haemochromatosis mapping to 7q22. Nature Genetics, 2000, 25(1): 14-15.

[28]Griffiths W J, Cox T M. Co-localization of the mammalian hemochromatosis gene product (HFE) and a newly identified transferrin receptor (TfR2) in intestinal tissue and cells. Journal of Histochemistry and Cytochemistry, 2003, 51(5): 613-624.

[29]Kawabata H, Fleming R E, Gui D, Moon S Y, Saitoh T, O’Kelly J, Umehara Y, Wano Y Said J W, Koeffler H P. Expression of hepcidin is down-regulated in TfR2 mutant mice manifesting a phenotype of hereditary hemochromatosis. Blood, 2005, 105(1): 376-381.

[30]Nemeth E, Roetto A, Garozzo G, Ganz T, Camaschella C. Hepcidin is decreased in TFR2 hemochromatosis. Blood, 2005, 105(4): 1803-1806.

[31]West A P Jr, Bennett M J, Sellers V M, Andrews N C, Enns C A, Bjorkman P J. Comparison of the interactions of transferrin receptor and transferrin receptor 2 with transferrin and the hereditary hemochromatosis protein HFE. Journal of Biological Chemistry, 2000, 275(49): 38135-38138.

[32]Robb A, Wessling-Resnick M. Regulation of transferrin receptor 2 protein levels by transferrin. Blood, 2004, 104(13): 4294-4299.

[33]Zhang M, Gumerov D R, Kaltashov I A, Mason A B. Indirect detection of protein-metal binding: interaction of serum transferrin with In3+ and Bi3+. Journal of the American Society for Mass Spectrometry, 2004, 15(11): 1658-1664.

[34]Evans G W. Transferrin function in zinc absorption and transport. Proceedings of the Society for Experimental Biology and Medicine, 1976, 151(4): 775-778.

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