Cloning and Tissue Expression Analysis of KLF3 Gene in Yak（Bos grunniens）

doi:10.13560/j.cnki.biotech.bull.1985.2017-0204

Abstract

Abstract: The aims of this study are to clone yak KLF3 gene sequence，to analyze its biological characterization，and to clarify its expression patterns in different tissues. Six healthy，4-6 years old male Maiwa yaks were selected as experiment animals. After fasting slaughter，the tissue samples of spleen，lung，kidney，subcutaneous fat and longissimus dorsi muscle were collected for the total RNA extraction. The Reverse Transcription PCR（RT-PCR）was used to clone Maiwa yak KLF3 gene sequence，the quantitative real-time PCR（qPCR）was used to detect the expression levels of KLF3 gene in different tissues. The results showed that the sequence of yak KLF3 gene was 1 137 bp（GenBank accession number：KX964630），containing 963 bp CDS，32 bp 5' UTR and 64 bp 3' UTR，encoding 346 amino acids. The amino acid sequence of yak KLF3 shared a similarity of 100% with that of cattle（XP_010820342.1）. The mRNA expression level of KLF3 was high in lung and liver tissue，significantly higher than other tissues（P < 0.01）.

Key words: yak, KLF3 Gene, clone, tissue expression

LIN Sen, LIN Ya-qiu, ZHU Jiang-jiang, BAI Xue, JIANG Ming-feng, WANG Yong. Cloning and Tissue Expression Analysis of KLF3 Gene in Yak（Bos grunniens）[J]. Biotechnology Bulletin, 2017, 33(8): 132-138.

References

[1] Pearson R, Fleetwood J, Eaton S, et al. Krüppel-like transcription factors：a functional family[J] . The International Journal of Biochemistry&Cell Biology, 2008, 40（10）：1996-2001.
[2] Chen Z, Lei T, Chen X, et al. Porcine KLF gene family：Structure, mapping, and phylogenetic analysis[J] . Genomics, 2010, 95（2）：111-119.
[3] Sue N, Jack BH, Eaton SA, et al. Targeted disruption of the basic Krüppel-like factor gene（Klf3）reveals a roal in adipogenesis[J] . Molecular and Cellular Biology, 2008, 28（12）：3967-3978.
[4] Turchinovich G, Vu TT, Frommer F, et al. Programming of marginal zone B-cell fate by basic Kruppel-like factor（BKLF/KLF3）[J] . Blood, 2011, 117（14）：529-537.
[5] Funnell AP, Maloney CA, Thompson LJ, at al. Erythroid Krüppel-like factor directly activates the basic Krüppel-like factor gene in erythroid cells[J] . Molecular and Cellular Biology, 2007, 27（7）：2777-2790.
[6] Himeda CL, Ranish JA, Pearson RC, at al. KLF3 regulates muscle-specific gene expression and synergizes with serum response factor on KLF binding sites[J] . Molecular and Cellular Biology, 2010, 30（14）：3430- 3443.
[7] Zhang J, Bakheet R, Parhar RS, et al. Regulation of fat storage and reproduction by Krüppel-like transcription factor KLF3 and fat-associated genes in Caenorhabditis elegans[J] . Journal of Molecular Biology, 2011, 411（3）：537-553.
[8] Zhang J, Hashmi S, Cheema F, et al. Regulation of lipoprotein assembly, secretion and fatty acid β-oxidation by Krüppel-like transcription factor, klf3[J] . Journal of Molecular Biology, 2013, 425（15）：2641-2655.
[9] Wu J, Srinivasan SV, Neumann JC, et al. The KLF2 transcription factor does not affect the formation of preadipocytes but inhibits their differentiation into adipocytes[J] . Biochemistry, 2005, 44（33）：11098-11105.
[10] Eaton SA, Funnell AP, Sue N, et al. A network of Kruppel-like factors（Klfs）, Klf8 is repressed by Klf3 and activated by Klf1 in vivo[J] . The Journal of Biological Chemistry, 2008, 283（40）：26937-26947.
[11] 万红玲, 雒林通, 吴建平. 牦牛肉品质特性研究进展[J] . 畜牧兽医杂志, 2012, 31（1）：36-40.
[12] 拜彬强, 郝力壮, 柴沙驼, 等. 牦牛肉品质特性研究进展[J] . 食品科学, 2014, 35（17）：290-296.
[13] 朱武政, 林亚秋, 江明锋, 等. 肉用山羊脂代谢相关基因与肌内脂肪含量的相关分析[J] . 畜牧兽医学报, 2016, 47（7）：1333-1341.
[14] Wang YH, Byrne KA, Reverter A, et al. Transcriptional profiling of skeletal muscle tissue from two breeds of cattle[J] . Mamm Genome, 2005, 16（3）：201-210.
[15] Zhang W, Kadam S, Emerson BM, et al. Site-specific acetylation by p300 or CREB binding protein regulates erythroid Kruppel-like factor transcriptional activity via its inetraction with the SWI-SNF complex[J] . Molecular and Cellular Biology, 2001, 21（7）：2431-2422.
[16] Kitamura T, Suzuki H, Tamura TA. Mouse Wee1 gene is repressed by Krüppel-like factor 3（KLF3）via interaction with multiple upstreamelements[J] . Gene, 2012, 492（2）：361-367.
[17] Dewi V, Kwok A, Lee S, et al. Phosphorylation of Krüppel-like factor 3（KLF3/BKLF）and C-terminal binding protein 2（CtBP2）by homeodomain-interacting protein kinase 2（HIPK2）modulates KLF3 DNA binding and activity[J] . The Journal of Biological Chemistry, 2015, 290（13）：8591-8605.
[18] 王茫桔, 瞿祥虎, 王立升, 等. 人类新型Krǜppel类转录因子hBKLF的cDNA克隆、亚细胞定位及表达特征[J] . 遗传学报, 2003, 30（1）：1-9.
[19] Crossley M, Whitelaw E, Perkins A, et al. Isolation and characterization of the cDNA encoding BKLF/TEF-2, a major CACCC-box-binding protein in erythroid cells and selected other cells[J] . Molecular and Cellular Biology, 1996, 16（4）：1695-1705.
[20] Zhang ZW, Wu CY, Li H, et al. Expression and functional analyses of Krüppel-like factor 3 in chicken adipose tissue[J] . Bioscience Biotechnolgy Biochemistry, 2014, 78（4）：614-623.
[21] 张志威, 李辉, 王宁. KLF转录因子家族与脂肪细胞分化[J] . 中国生物化学与分子生物学报, 2009, 25（11）：983-990.