Establishment of interferon membrane protein IFITML knockdown cells and investigation of its effect on the replication of PRV, PCV2 and TGEV
Chinese Veterinary Science / Zhongguo Shouyi Kexue
; 50(5):603-609, 2020.
Article
in Chinese
| CAB Abstracts | ID: covidwho-1994656
ABSTRACT
To establish a J2-KD (knockdown) cell line stably expressing interfered IFITM1 and study the effect of interference with IFITMI gene on the infection of PCV2, PRV and TGEV. Gene cloning tech- niques were used to constructed pLKO. l-EGFP-Puro-IFITMI recombinant vector, which was co-transfected into 293 FT cells with lentiviral packaging plasmids psPAXZ and pMDZ. G to produce green fluorescent protein labeled lentiviruses expression IFITMlshRNA, the viral supernatant was collected at 48 hours after post transfection. J2 cells were infected with the harvested lentiviruses, screened by puromycin and cloned via cell limited dilution. Real-time PCR identify that the cell lines with stable interference with IFITMl gene were obtained, and via MTT method verify that interference with IFITMI expression had no effect on the growth of J2 cells, the successfully constructed J2 stable cell line interfere with IFITMl expression was named as JZ-KD. PRV, PCV2 and TGEV infected J2-KD cells, respectively. Using real-time fluorescence quantitative PCR detect virus replication. The results showed that J2-KD cell line was successfully generated with interfered IFITMl expression;the copy number of PCV2 and TGEV were in- creased, while PRV was decreased in J'Z-KD cell. Indicating that the interference of IFITMI gene expression markedly inhibited the replication of PRV while promoted that. of TGEV and PCV2, providing a basis for further study on the function of porcine IFITMI protein and elucidates its antiviral mechanism.
Prion, Viral, Bacterial and Fungal Pathogens of Animals [LL821]; Diagnosis of Animal Diseases [LL886]; Genetic Engineering, Gene Transfer and Transgenics [WW100]; Animal Immunology [LL650]; Host Resistance and Immunity [HH600]; cell lines; gene expression; viral replication; genes; interferons; swine diseases; viral diseases; DNA cloning; antiviral agents; Suid herpesvirus 1; Transmissible gastroenteritis virus; pigs; Porcine circovirus 2; Varicellovirus; Alphaherpesvirinae; Herpesviridae; Herpesvirales; dsDNA Viruses; DNA Viruses; viruses; Alphacoronavirus 1; Alphacoronavirus; Coronavirinae; Coronaviridae; Nidovirales; positive-sense ssRNA Viruses; ssRNA Viruses; RNA Viruses; Sus scrofa; Sus; Suidae; Suiformes; Artiodactyla; mammals; vertebrates; Chordata; animals; eukaryotes; Circovirus; Circoviridae; ssDNA Viruses; Aujeszky virus; pig diseases; viral infections; swine; hogs; antivirals
Full text:
Available
Collection:
Databases of international organizations
Database:
CAB Abstracts
Type of study:
Experimental Studies
Language:
Chinese
Journal:
Zhongguo Shouyi Kexue
Year:
2020
Document Type:
Article
Similar
MEDLINE
...
LILACS
LIS