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Molecular characterization of Sp110 gene in pigs

Author:
Li, Li-zu, Wang, Qiu-shi, Han, Li-xin, Wang, Jin-kui, Shao, Si-yu, Wang, Liang, Liu, Di, Yang, Xiu-qin
Source:
Molecular genetics and genomics 2016 v.291 no.3 pp. 1431-1442
ISSN:
1617-4615
Subject:
alternative splicing, double-stranded RNA, genes, half life, humans, infectious diseases, livestock breeding, quantitative polymerase chain reaction, reverse transcriptase polymerase chain reaction, reverse transcription, swine
Abstract:
Speckled 110 kDa (Sp110) plays an important role in infectious diseases, as revealed by studies in humans. However, little is known regarding porcine Sp110. To elucidate its potential role in porcine resistance to viral diseases, here, the complete coding sequence of porcine Sp110 gene and its 26 alternatively spliced isoforms were isolated using reverse transcription (RT)-polymerase chain reaction (PCR), and another seven splicing patterns were obtained using a minigene construct. Subcellular distribution of 11 representative isoforms was characterized in PK-15 cells transiently transfected with their respective GFP fusion constructs, and only isoforms (R and V) bearing all functional domains were localized in nucleus, indicating all the other isoforms lose normal functions of Sp110 owing to alternative splicing. Real-time quantitative PCR and competitive RT-PCR showed that both isoforms R and V had similar tissue expression profile, half-life and response to poly(I:C), a synthetic analog of viral double-stranded RNA, while the longer one (isoform R) was transcribed at a higher level. The results indicated that porcine Sp110 has a role in viral infection and that isoform R is the dominant active form. Overall the data provide potential resource for molecular breeding of pig resistant to diseases and contributes to breeding pigs resistant to viral infection.
Agid:
5207469