Partial characterization of three &#946-defensin gene transcripts in river buffalo and cattle

In this study, the tracheal tissues from Egyptian river buffalo and cattle were screened for the presence of three bovine β-defensin gene transcripts. Three primer pairs were designed on the basis of published Bos taurus sequences for partial amplification of β-defensin 4, β-defensin 10 and β-defensin 11 complementary DNA (cDNA). The amplified cDNA products of the three genes in both buffalo and cattle were sequenced. The sequences were analyzed to verify gene identity and to identify differences with the corresponding buffalo (Bubalus bubalis bubalis) and/or cattle (Bos taurus) β-defensin mRNAs published sequences in the GenBank database. β-Defensin 4 and β-defensin 10 primer pairs amplified cDNA sequences in buffalo and cattle that corresponded to those mRNAs of the two genes in GenBank database with nucleotide percentage homology of 83 and 84% for β-defensin 4, and 87 and 90% for β-defensin10, respectively. The translated protein sequences obtained for buffalo and cattle showed protein percentage similarity of 86 and 81% for β-defensin 4, and 87.5 and 87% for β-defensin 10 with the corresponding proteins of B. bubalis bubalis and/or B. taurus in GenBank database. On the other hand, cDNA sequences amplified by β-defensin 11 primer pair in both buffalo and cattle corresponded more to lingual antimicrobial peptide (LAP) mRNAs of B. bubalis bubalis and B. taurus (94 and 82% nucleotide similarity and 92 and 77% translated-protein similarity) rather than β-defensin 11 mRNA of B. taurus (68 and 66% nucleotide similarity and 74 and 65.5% translated-protein similarity).Key words: β-Defensin 4, β-defensin 10, β-defensin 11, lingual antimicrobial peptide (LAP), river buffalo, cattle

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

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.Key words: CSRP3, cattle, river buffalo

Analysis of genetic variation in different sheep breeds using microsatellites

Genetic variation in three Egyptian indigenous sheep breeds namely: Barki, Ossimi and Rahmani were investigated using fourteen microsatellite loci. The total number of alleles ranged from 6 in CSSM47 locus to 14 in TGLA 377 locus. The fourteen tested loci were all polymorphic in the three breeds. Major differences between the breeds were found at ten of the tested loci, where the alleles at the highest frequency are different in the three breeds. While, at loci OARCP20, OARVH72, CSSM47 and OARAE129, two of the tested breeds have similar alleles at the highest allele frequency. The average direct count of heterozygosity overall loci in each tested breed was less than the expected heterozygosity. Tests of genotype frequencies for deviation from the Hardy-Weinberg equilibrium (HWE), at each locus overall breeds, revealed significant departure from HWE due to heterozygote deficiency. A slightly high rate of inbreeding within the three breeds was noticed (global FIS = 0.308). Low genetic differentiation was detected by estimation of FST index between all pairs of breeds. Cluster analysis revealed that Ossimi and Rahmani breeds clustered independently from Barki breed at 0.43 of genetic distance. The obtained results can be useful for the development of a rational breeding strategy for genetic improvement of sheep in Egypt