Characterization and sequence analysis of cysteine and glycine-rich protein 3 in Egyptian native cattle and river native buffalo cDNA sequences

Abstract

Cysteine and glycine rich protein, CSRP3 also referred to as the muscle LIM protein (MLP), has been investigated in native Egyptian cattle and buffalo (river buffalo). RNA extraction and cDNA synthesis were conducted from different tissue samples. Primers specific for CSRP3 were designed using known cDNA sequences of Bos taurus published in database with different accession numbers. Polymerase chain reaction (PCR) was performed and products were purified and sequenced. Sequence analysis and alignment were carried out using CLUSTAL W (1.83). Multiple nucleotide sequence alignment between CSRP3 cDNA amplicons of native buffalo and cattle revealed 89% identity. B. taurus CSRP3 mRNA (Cardiac LIM protein) [NM 001024689.2] showed 85 and 87% identity in nucleic acid sequences and 82 and 84% homology in amino acid sequences with native cattle and buffalo, respectively. A 90% homology was detected between the amino acid sequences of river buffalo and native cattle. Fourty nine translated amino acids out of 51 in both buffalo and cattle are found to be part of the conserved CSRP3 LIM1 domain protein which comprises 57 codons. The LIM1 domain in Egyptian buffalo and cattle CSRP3 showed only 87 and 85% similarity with B. taurus CSRP3 LIM1 domain, respectively, which are caused mainly by frame shift mutation resulting from a single nucleotide deletion. Sequence nucleotide alignment of both native buffalo and cattle CSRP3 cDNAs sequences and B. taurus whole genome showed high percent identity (94-100%) with B. taurus chromosome 29 |NC-007330.3|. This confirmed the assignment of CSRP3 to cattle chromosome BTA 29 and allowed the indirect assignment of CSRP3 to river buffalo chromosome BBU5p (the homologue of BTA 29) based on the extensive chromosome homology and conservation between cattle and river buffalo.