CDNA Cloning, Sequence Identification and Tissue Expression Distribution of Three Novel Genes: DFP, ITM2B and PQLC1 from Black-Boned Sheep (Ovis Aries)

ABSTRACT

The complete coding sequences of three of Black-boned sheep (Ovis aries) genes DFP, ITM2B and PQLC1 were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) according to the conserved sequence information of the cattle or other mammals and known highly homologous sheep ESTs. Black-boned sheep DFP gene encodes a protein of 213 amino acids which contains the conserved putative DNA/pantothenate metabolism flavoprotein domain and has high homology with the DFP proteins of four species: cattle (99%), dog (91%), human (88%) and zebrafish (53%). Black-boned sheep ITM2B gene encodes a protein of 266 amino acids that contains the conserved putative BRICHOS domain and has high similarity with the ITM2B proteins of five species: cattle (98%), human (95%), mouse (94%), rat (94%) and pig (94%). Black-boned sheep PQLC1 gene encodes a protein of 253 amino acids that contains the conserved putative PQ-loop repeat domain and has high homology with the PQLC1 proteins of four species: cattle (99%), human (91%), mouse (90%) and rat (84%). The phylogenetic tree analysis demonstrated that Black-boned sheep's DFP, ITM2B andPQLC1 proteins have a close relationship with cattle. The tissue expression analysis indicated that Black-boned sheep DFP, ITM2B and PQLC1 genes were expressed in a range of tissues including leg muscle, kidney, skin, longissimus dorsi muscle, spleen, heart and liver. Our experiment is the first to provide the primary foundation for further insight into these three sheep genes.

Keywords:

• Black-boned sheep
• DFP
• ITM2Band PQLC1
• tissue expression analysis