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| Cloning and Expression Analysis of Goat (Capra hircus) KLF13 Gene |
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Abstract Krüppel-like factors (KLFs) is a group of zinc finger protein which is important for gene transcriptional regulation in eucaryote. The aim of the study was to clone the CDS of KLF13 gene, and to determine the abundance of KLF13 mRNA expression in various tissues and differential stages of preadipocytes, which may provide the basic information about the fuction of KLF13 gene on regulating preadipocytes differentiation. Four 24-month-old healthy Jianzhou Big-Eared male goats (Capra hircus) were selected randomly in this study. After slaughtered, the tissue samples of heart, liver, spleen, lungs, kidney, subcutaneous fat, longissimus dosi muscle, biceps femoris muscle and triceps brachii muscle were collected for the total RNA extraction. The sequence of KLF13 gene was cloned by RT-PCR and analyzed using bioinformatics. The mRNA expression of KLF13 in various tissues, preadipocytes (0, 3, 5, 7 d) and myoblasts (0, 2, 3, 4, 5 d) were determined using qRT-PCR. A length of 991 bp cDNA of KLF13 gene was cloned (GenBank No.: KU041755), which contains 876 bp CDS, 12 bp 5' UTR and 103 bp 3' UTR, encoding 292 amino acids. The goat KLF13 protein shared 100%、97.9%、96.6% and 90.8% similarity with that of sheep (Ovis aries), bovine (Bos taurus), pig (Sus scrofa) and mouse (Mus musculus), respectively. Goat KLF13 mRNA sequence was closest to that of sheep, following by bovine, human (Homo sapiens), pig and mouse. The result of tissue expression showed that KLF13 gene was expressed high in spleen and lungs, and low in muscle. The expression of KLF13 was continually suppressed significantly with the differentiation of goat preadipocytes. However, excepted at 5 d, the expression of KLF13 was enhanced with the differentiation of myoblasts. These results showed that KLF13 gene may be involved in preadipocytes differentiation, and also will be beneficial for studying the role of KLF13 in regulating preadipocytes differentiation in goat.
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Received: 14 September 2016
Published: 24 December 2016
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