molecularcloning,sequencecharacterization,polymorphism.docVIP

Molecular cloning, sequence characterization, polymorphism and association analysis of porcine ROPN1 gene Lan Jing1, Zhao Jiugang1, Liu Yonggang2 1Chongqing Key Laboratory of Pig Industry Sciences, Chongqing Academy of Animal Sciences, Rongchang 402460, China 2College of Animal Science and Technology, Yunnan Agricultural University, Kunming 650201, China Abstract The full-length cDNA sequence of one porcine gene, ROPN1, was isolated using the rapid amplification of cDNA ends (RACE) method based on one pig EST sequence which was highly homologous to the coding sequence of human ROPN1 gene. The porcine ROPN1 gene encodes a protein of 212 amino acids which shares high homology with the rhophilin associated protein 1 (ROPN1) of eight species: gray short-tailed opossum (96%), horse (95%), cattle (94%), mouse (93%), rat (92%), chimpanzee (85%), human (85%) and rhesus monkey (85%). Phylogenetic analysis revealed that the porcine ROPN1 gene has a closer genetic relationship with the ROPN1 gene of gray short-tailed opossum. Polymorphism analysis showed that there was a T/C mutation at the position of 536-bp of mRNA and this leaded to the amino acid alteration from the Arg residue to the Cys residue. PCR- Hae III -RFLP was established to detect this T/C mutation and eight pig breeds display obvious genotype and allele frequency differences at this mutation locus. Association of this SNP with litter size traits was assessed in Large White (n=100) and Landrace (n=100) pig populations, and results demonstrated that this polymorphic locus was significantly associated with the litter size of first parity (P0.01) and all parities (P0.05) in Large White sows, and also significantly associated with the litter size of all parities in Landrace sows (P0.01). Therefore, ROPN1 gene could be a useful candidate gene in selection for increasing litter size in pigs. These data serve as a foundation for further insight into this novel porcine gene. Key words: pig, ROPN1, polymorphism, asso