ISOLATION AND SEQUENCING OF MRF4 GENE (EXON 1 THROUGH 4) FROM EGYPTIAN BUFFALO

Document Type : Original Article

Authors

1 Department of biochemistry, Fac. of Vet. Med. Kafrelsheikh Univ., Egypt.

2 Department of clinical pathology, Fac. of Vet. Med. Kafrelsheikh Univ., Egypt.

3 Department of Anatomy and Embryology, Fac. of Vet. Med. Kafrelsheikh Univ., Egypt.

Abstract

Muscle regulatory factor 4(MRF4, also known as herculin and MYF6) is a member of MyoD family, which is involved in the differentiation and maturation of myotubes and highly expressed postnatally.The sequences of this gene are well known in cattle and small ruminants but in buffalo no available sequence was previously recorded before this research. Therefore, the main aim of this study is to isolate and sequence exon 1 (E1) through exon 4 (E4) portion of this gene, which is the most appropriate portion of MRF4 that contains single nucleotide polymorphisms (SNPs), in buffalo. To achieve this, RT-PCR was performed in which gene-specific primers flanking to E1-E4 of MRF4 were utilized in addition to PCR master mix. A PCR product of 693bp, of MRF4 gene corresponding to the expected theoretical product size, was successfully amplified. Sequencing of the purified PCR product and its alignment against a previously known bovine gene identified 4 synonymous novel SNPs in buffalo MRF4. This result indicates that the sequence of MRF4 is highly conserved between cattle and buffalo. This is a preliminary study that provides the researchers withraw data which could be used as a basis for further studies to associate these SNPs with meat quality traits in buffalo.

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Kafrelsheikh Veterinary Medical Journal
Volume 10, Issue 2
October 2012
Pages 57-67

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