Cloning the Horse RNA Polymerase I Promoter and Its Application to Studying Influenza Virus Polymerase Activity.
An influenza virus polymerase reconstitution assay based on the human, dog, or chicken RNA polymerase I (PolI) promoter has been developed and widely used to study the polymerase activity of the influenza virus in corresponding cell types. Although it is an important member of the influenza virus family and has been known for sixty years, no studies have been performed to clone the horse PolI promoter or to study the polymerase activity of equine influenza virus (EIV) in horse cells. In our study, the horse RNA PolI promoter was cloned from fetal equine lung cells. Using the luciferase assay, it was found that a 500 bp horse RNA PolI promoter sequence was required for efficient transcription. Then, using the developed polymerase reconstitution assay based on the horse RNA PolI promoter, the polymerase activity of two EIV strains was compared, and equine myxovirus resistance A protein was identified as having the inhibiting EIV polymerase activity function in horse cells. Our study enriches our knowledge of the RNA PolI promoter of eukaryotic species and provides a useful tool for the study of influenza virus polymerase activity in horse cells.
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. 18814113742@163.com.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. 18814113742@163.com.
Gang
G
Lu
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. donghe9108@163.com.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. donghe9108@163.com.
Dong
D
He
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. lugang19860501@163.com.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. lugang19860501@163.com.
Zengchao
Z
Wang
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. whj751364486@163.com.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. whj751364486@163.com.
Shudan
S
Ou
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. jeremyjessica@163.com.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. jeremyjessica@163.com.
Rong
R
Yuan
College of Veterinary Medicine, South China Agricultural University, Guangzhou 510000, China. shoujunli@scau.edu.cn.|Key Laboratory of Comprehensive Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province, Guangzhou 510000, China. shoujunli@scau.edu.cn.
Shoujun
S
Li
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism*; D006736:Horses; D009980:Influenza A virus / Q000201:enzymology* / Q000502:physiology; D011401:Promoter Regions, Genetic; D012318:RNA Polymerase I / Q000235:genetics*; D014158:Transcription, Genetic; D014779:Virus Replication
D012321:DNA-Directed RNA Polymerases; D012318:RNA Polymerase I
RNA polymerase I promoter; equine influenza virus; polymerase activity