pAPN基因敲除的IPEC-J2介导的TGEV感染特征分析

doi:10.11843/j.issn.0366-6964.2024.08.013

Abstract

Abstract:

The aim of this study was to explore the characteristics of porcine transmissible gastroenteritis virus (TGEV) infection mediated by porcine aminopeptidase N (pAPN) gene knockout intestinal porcine epithelial cell line J2 (IPEC-J2) and to provide theoretical basis for further understanding the mechanism of pAPN gene in the process of TGEV infection. The study was divided into 3 groups, pAPN gene knockout IPEC-J2 group (IPEC-J2-KO group), wild-type IPEC-J2 group (IPEC-J2-WT group), and wild-type IPEC-J2 group not inoculated with TGEV (Mock group), with 3 replicates set up in each group. Firstly, quantitative real-time PCR (qPCR) was used to determine the best time point for collecting cell samples following inoculation with TGEV strain. Secondly, the off-target effect of IPEC-J2-KO was detected. Then, the infection characteristics of IPEC-J2-KO, IPEC-J2-WT inoculated with TGEV and Mock were analyzed by qPCR, western blot (WB), indirect immunofluorescence assay (IFA), and 50% tissue culture infective dose (TCID₅₀). Finally, the expression of NF-κB p65 and its phosphorylated protein pp65 in Mock, IPEC-J2-WT and IPEC-J2-KO were detected by WB. The results of qPCR showed that 24 hours after exposure was the best time point to collect cell samples to evaluate the impact of TGEV on IPEC-J2; The off-target analysis results showed that no off-target effects were detected in IPEC-J2-KO; The results of virus infection characteristic analysis showed that the virus copy number and virus titer in IPEC-J2-KO were significantly lower than those in IPEC-J2-WT (P < 0.001). Compared with the Mock, there was no significant difference in virus copy number and virus titer in IPEC-J2-KO (P>0.05), and no expression of TGEV-N protein was detected in IPEC-J2-KO. In addition, compared with the Mock, after inoculation with TGEV, the phosphorylation level of NF-κB p65 was significantly increased in the IPEC-J2-WT group (P < 0.001), while there was no significant difference in IPEC-J2-KO group (P>0.05). This study results showed that IPEC-J2-KO can effectively resist TGEV infection, and TGEV did not affect the activity of the transcription factor NF-κB in the innate immunity-related signaling pathway in IPEC-J2-KO. This study provided the evidence that IPEC-J2 could serve as a cell model for the study of TGEV infection characteristics, and laid the foundation for elucidating the mechanism of pAPN gene in TGEV invading host cells and researching new disease-resistant pig varieties.

Key words: porcine aminopeptidase N, IPEC-J2, TGEV, NF-κB

CLC Number:

S813.1

Zhentao XIA, Nan WANG, Wanjie WANG, Qilü ZHOU, Lei HUANG, Yulian MU. Characteristics Analysis of TGEV Infection Mediated by IPEC-J2 with Knockout of pAPN Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(8): 3395-3407.

Figures/Tables 9

Table 1

Table 2

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

References 58

1	LIU Q , WANG H Y . Porcine enteric coronaviruses: an updated overview of the pathogenesis, prevalence, and diagnosis[J]. Vet Res Commun, 2021, 45 (2-3): 75- 86. doi: 10.1007/s11259-021-09808-0
2	DOYLE L P , HUTCHINGS L M . A transmissible gastroenteritis in pigs[J]. J Am Vet Med Assoc, 1946, 108, 257- 259.
3	CHEN S Y , ZHANG H Y , CHU M F , et al. Prevalence of transmissible gastroenteritis among swine populations in China during 1983-2022: A systematic review and meta-analysis[J]. Microb Pathog, 2023, 183, 106320. doi: 10.1016/j.micpath.2023.106320
4	TURLEWICZ-PODBIELSKA H , POMORSKA-MÓL M . Porcine Coronaviruses: Overview of the State of the Art[J]. Virol Sin, 2021, 36 (5): 833- 851. doi: 10.1007/s12250-021-00364-0
5	曹贝贝, 兰培英, 韩丽, 等. 猪TGEV HN-2012株ORF3a和ORF3b基因遗传变异分析及其真核表达研究[J]. 西北农林科技大学学报(自然科学版), 2016, 44 (4): 10- 16.
	CAO B B , LAN P Y , HAN L , et al. Genetic variation and eukaryotic expression of ORF3a and ORF3b from porcine TGEV HN-2012 strain[J]. Journal of Northwest A&F University (Natural Science Edition), 2016, 44 (4): 10- 16.
6	郑宾宾, 宋涛, 季裕婷, 等. 猪传染性胃肠炎病毒HB-1株的分离鉴定和全基因组序列分析[J]. 中国兽医杂志, 2023, 59 (5): 36- 41.
	ZHENG B B , SONG T , JI Y T , et al. Isolation, identification and whole genome sequence analysis of porcine transmissible gastroenteritis virus HB-1 strain[J]. Chinese Journal of Veterinary Medicine, 2023, 59 (5): 36- 41.
7	李嘉琛, 吴华伟, 陈晓春, 等. 猪传染性胃肠炎病毒的分离与鉴定[J]. 中国兽药杂志, 2018, 52 (3): 25- 30.
	LI J C , WU H W , CHEN X C , et al. Isolation and identification of porcine transmissible gastroenteritis virus[J]. Chinese Journal of Veterinary Drug, 2018, 52 (3): 25- 30.
8	俞伏松, 王劭, 陈仕龙, 等. 猪传染性胃肠炎病毒福建株的分离鉴定[J]. 福建农业学报, 2012, 27 (11): 1160- 1164. doi: 10.3969/j.issn.1008-0384.2012.11.003
	YU F S , WANG S , CHEN S L , et al. Isolation and identification of porcine transmissible gastroenteritis virus FJ strain[J]. Fujian Journal of Agricultural Sciences, 2012, 27 (11): 1160- 1164. doi: 10.3969/j.issn.1008-0384.2012.11.003
9	祖立闯, 王芳, 刘爱华, 等. 我国猪传染性胃肠炎的流行现状及实验室诊断技术研究进展[J]. 养猪, 2017, (5): 123- 128. doi: 10.3969/j.issn.1002-1957.2017.05.041
	ZU L C , WANG F , LIU A H , et al. Research progress of epidemic status and laboratory diagnostic technology of porcine transmissible gastroenteritis in China[J]. Swine Production, 2017, (5): 123- 128. doi: 10.3969/j.issn.1002-1957.2017.05.041
10	YIN L , LIU X , HU D M , et al. Swine enteric coronaviruses (PEDV, TGEV, and PDCoV) induce divergent interferon-stimulated gene responses and antigen presentation in porcine intestinal enteroids[J]. Front Immunol, 2021, 12, 826882.
11	VLASOVA A N , DIAZ A , DAMTIE D , et al. Novel canine coronavirus isolated from a hospitalized patient with pneumonia in east malaysia[J]. Clin Infect Dis, 2022, 74 (3): 446- 454. doi: 10.1093/cid/ciab456
12	CHEN Y W , ZHANG Y Z , WANG X , et al. Transmissible gastroenteritis virus: An update review and perspective[J]. Viruses, 2023, 15 (2): 359. doi: 10.3390/v15020359
13	ALONSO S , IZETA A , SOLA I , et al. Transcription regulatory sequences and mRNA expression levels in the coronavirus transmissible gastroenteritis virus[J]. J Virol, 2002, 76 (3): 1293- 1308. doi: 10.1128/JVI.76.3.1293-1308.2002
14	GALÁN C , SOLA I , NOGALES A , et al. Host cell proteins interacting with the 3' end of TGEV coronavirus genome influence virus replication[J]. Virology, 2009, 391 (2): 304- 314. doi: 10.1016/j.virol.2009.06.006
15	REN X F , LIU B Q , YIN J C , et al. Phage displayed peptides recognizing porcine aminopeptidase N inhibit transmissible gastroenteritis coronavirus infection in vitro[J]. Virology, 2011, 410 (2): 299- 306. doi: 10.1016/j.virol.2010.11.014
16	RIEMANN D , KEHLEN A , LANGNER J . CD13-not just a marker in leukemia typing[J]. Immunol Today, 1999, 20 (2): 83- 88. doi: 10.1016/S0167-5699(98)01398-X
17	DELMAS B , GELFI J , KUT E , et al. Determinants essential for the transmissible gastroenteritis virus-receptor interaction reside within a domain of aminopeptidase-N that is distinct from the enzymatic site[J]. J Virol, 1994, 68 (8): 5216- 5224. doi: 10.1128/jvi.68.8.5216-5224.1994
18	YEAGER C L , ASHMUN R A , WILLIAMS R K , et al. Human aminopeptidase N is a receptor for human coronavirus 229E[J]. Nature, 1992, 357 (6377): 420- 422. doi: 10.1038/357420a0
19	SODERBERG C , GIUGNI T D , ZAIA J A , et al. CD13 (human aminopeptidase N) mediates human cytomegalovirus infection[J]. J Virol, 1993, 67 (11): 6576- 6585. doi: 10.1128/jvi.67.11.6576-6585.1993
20	DELMAS B , GELFI J , L'HARIDON R , et al. Aminopeptidase N is a major receptor for the enteropathogenic coronavirus TGEV[J]. Nature, 1992, 357 (6377): 417- 420. doi: 10.1038/357417a0
21	LI W T , LUO R , HE Q G , et al. Aminopeptidase N is not required for porcine epidemic diarrhea virus cell entry[J]. Virus Res, 2017, 235, 6- 13. doi: 10.1016/j.virusres.2017.03.018
22	WANG B , LIU Y , JI C M , et al. Porcine deltacoronavirus engages the transmissible gastroenteritis virus functional receptor porcine aminopeptidase N for infectious cellular entry[J]. J Virol, 2018, 92 (12): e00318- 18.
23	ZHU X Y , LIU S D , WANG X L , et al. Contribution of porcine aminopeptidase N to porcine deltacoronavirus infection[J]. Emerg Microbes Infect, 2018, 7 (1): 65.
24	XU K , ZHOU Y R , MU Y L , et al. CD163 and pAPN double-knockout pigs are resistant to PRRSV and TGEV and exhibit decreased susceptibility to PDCoV while maintaining normal production performance[J]. Elife, 2020, 9, e57132. doi: 10.7554/eLife.57132
25	DEDIEGO M L , NIETO-TORRES J L , JIMENEZ-GUARDENO J M , et al. Coronavirus virulence genes with main focus on SARS-CoV envelope gene[J]. Virus Res, 2014, 194, 124- 137. doi: 10.1016/j.virusres.2014.07.024
26	GUO J X , ZHAO X M , LIU Z H , et al. Transmissible gastroenteritis virus ORF3b up-regulates miR-885-3p to counteract TNF-alpha production via inhibiting NF-kappaB pathway[J]. Vet Microbiol, 2021, 261, 109189. doi: 10.1016/j.vetmic.2021.109189
27	WANG L , QIAO X Y , ZHANG S J , et al. Porcine transmissible gastroenteritis virus nonstructural protein 2 contributes to inflammation via NF-kappaB activation[J]. Virulence, 2018, 9 (1): 1685- 1698. doi: 10.1080/21505594.2018.1536632
28	ZHOU Y R , WU W , XIE L L , et al. Cellular RNA helicase DDX1 is involved in transmissible gastroenteritis virus nsp14-induced interferon-beta production[J]. Front Immunol, 2017, 8, 940. doi: 10.3389/fimmu.2017.00940
29	WU Y , LI M W , TIAN J , et al. Broad antagonism of coronaviruses nsp5 to evade the host antiviral responses by cleaving POLDIP3[J]. PLoS Pathog, 2023, 19 (10): e1011702. doi: 10.1371/journal.ppat.1011702
30	DU J , CHEN D W , YU B , et al. L-Leucine promotes STAT1 and ISGs expression in TGEV-infected IPEC-J2 cells via mTOR activation[J]. Front Immunol, 2021, 12, 656573. doi: 10.3389/fimmu.2021.656573
31	GUO Z Z , ZHANG C X , DONG J J , et al. Persistence infection of TGEV promotes enterococcus faecalis infection on IPEC-J2 cells[J]. Int J Mol Sci, 2022, 24 (1): 450. doi: 10.3390/ijms24010450
32	王晓朋, 徐奎, 魏迎辉, 等. CRISPR/Cas9介导的猪IPEC-J2细胞CD13基因敲除细胞系的建立[J]. 畜牧兽医学报, 2019, 50 (7): 1319- 1327.
	WANG X P , XU K , WEI Y H , et al. Establishment of CD13 gene knockout IPEC-J2 cell lines mediated by CRISPR/Cas9 system[J]. Acta Veterinaria et Zootechnica Sinica, 2019, 50 (7): 1319- 1327.
33	GUO R L , FAN B C , CHANG X J , et al. Characterization and evaluation of the pathogenicity of a natural recombinant transmissible gastroenteritis virus in China[J]. Virology, 2020, 545, 24- 32. doi: 10.1016/j.virol.2020.03.001
34	XIA L , YANG Y H , WANG J L , et al. Impact of TGEV infection on the pig small intestine[J]. Virol J, 2018, 15 (1): 102. doi: 10.1186/s12985-018-1012-9
35	GUO J Y , LI F , QIAN S J , et al. TGEV infection up-regulates FcRn expression via activation of NF-kappaB signaling[J]. Sci Rep, 2016, 6, 32154. doi: 10.1038/srep32154
36	ZHAO X M , MA X L , GUO J X , et al. Circular RNA CircEZH2 suppresses transmissible gastroenteritis coronavirus-induced opening of mitochondrial permeability transition pore via targeting MiR-22 in IPEC-J2[J]. Int J Biol Sci, 2019, 15 (10): 2051- 2064. doi: 10.7150/ijbs.36532
37	PFLEIDERER G , CELLIERS P G . Isolation of an aminopetidase from kidney particles[J]. Biochem Z, 1963, 339, 186- 189.
38	KUMARAVEL S , LUO G R , HUANG S T , et al. Development of a novel latent electrochemical molecular substrate for the real-time monitoring of the tumor marker aminopeptidase N in live cells, whole blood and urine[J]. Biosens Bioelectron, 2022, 203, 114049. doi: 10.1016/j.bios.2022.114049
39	CHEN L , LIN Y L , PENG G Q , et al. Structural basis for multifunctional roles of mammalian aminopeptidase N[J]. Proc Natl Acad Sci U S A, 2012, 109 (44): 17966- 17971. doi: 10.1073/pnas.1210123109
40	WHITWORTH K M , ROWLAND R R , EWEN C L , et al. Gene-edited pigs are protected from porcine reproductive and respiratory syndrome virus[J]. Nat Biotechnol, 2016, 34 (1): 20- 22. doi: 10.1038/nbt.3434
41	SHI X , TANG T , LIN Q Y , et al. Efficient generation of bone morphogenetic protein 15-edited Yorkshire pigs using CRISPR/Cas9?[J]. Biol Reprod, 2020, 103 (5): 1054- 1068. doi: 10.1093/biolre/ioaa138
42	许美娜, 朱奕舟, 林思远, 等. CRISPR/Cas9基因编辑技术在猪育种中的研究进展[J]. 广东农业科学, 2022, 49 (8): 87- 96.
	XU M N , ZHU Y Z , LIN S Y , et al. Progress of the application of CRISPR/Cas9 gene editing technology in pig breeding[J]. Guangdong Agricultural Sciences, 2022, 49 (8): 87- 96.
43	XIANG G H , REN J L , HAI T , et al. Editing porcine IGF2 regulatory element improved meat production in Chinese Bama pigs[J]. Cell Mol Life Sci, 2018, 75 (24): 4619- 4628. doi: 10.1007/s00018-018-2917-6
44	ZHENG Q T , LIN J , HUANG J J , et al. Reconstitution of UCP1 using CRISPR/Cas9 in the white adipose tissue of pigs decreases fat deposition and improves thermogenic capacity[J]. Proc Natl Acad Sci U S A, 2017, 114 (45): E9474- E9482.
45	XU K , ZHOU Y R , SHANG H T , et al. Pig macrophages with site-specific edited CD163 decrease the susceptibility to infection with porcine reproductive and respiratory syndrome virus[J]. J. Integr. Agric, 2023, 22 (7): 2188- 2199. doi: 10.1016/j.jia.2022.11.010
46	NIU D , WEI H J , LIN L , et al. Inactivation of porcine endogenous retrovirus in pigs using CRISPR-Cas9[J]. Science, 2017, 357 (6357): 1303- 1307. doi: 10.1126/science.aan4187
47	MOTOCHE-MONAR C , ORDONEZ J E , CHANG O , et al. gRNA Design: How its evolution impacted on CRISPR/Cas9 systems refinement[J]. Biomolecules, 2023, 13 (12): 1698. doi: 10.3390/biom13121698
48	ZHANG S J , HUANG W Z , REN L L , et al. Comparison of viral RNA-host protein interactomes across pathogenic RNA viruses informs rapid antiviral drug discovery for SARS-CoV-2[J]. Cell Res, 2022, 32 (1): 9- 23. doi: 10.1038/s41422-021-00581-y
49	张羽欣, 王树茂, 段宏勇, 等. 猪传染性胃肠炎病毒TaqMan实时荧光定量PCR检测方法的建立与应用[J]. 中国兽医科学, 2024, (4): 1- 8.
	ZHANG Y X , WANG S M , DUAN H Y , et al. Establishment and application of TaqMan real-time quantitative PCR for detection of porcine transmissible gastroenteritis virus[J]. Chinese Veterinary Science, 2024, (4): 1- 8.
50	董珮玲. 猪传染性胃肠炎病毒N蛋白单克隆抗体的制备及其初步应用[D]. 武汉: 华中农业大学, 2023.
	DONG P L. Preparation of monoclonal antibody to Transmissiblegastroenteritis virus N protein and its initial application[D]. Wuhan: Huazhong Agricultural University, 2023. (in Chinese)
51	WHITWORTH K M , ROWLAND R , PETROVAN V , et al. Resistance to coronavirus infection in amino peptidase N-deficient pigs[J]. Transgenic Res, 2019, 28 (1): 21- 32. doi: 10.1007/s11248-018-0100-3
52	XIAO W W , WANG X L , WANG J , et al. Replicative capacity of four porcine enteric coronaviruses in LLC-PK1 cells[J]. Arch Virol, 2021, 166 (3): 935- 941. doi: 10.1007/s00705-020-04947-2
53	ZHANG S S , CAO Y N , XU C , et al. Integrated metabolomics and transcriptomics analyses reveal metabolic responses to TGEV infection in porcineintestinal epithelial cells[J]. J Gen Virol, 2023, 104 (12): 10.
54	MA X L , ZHAO X M , ZHANG Z C , et al. Differentially expressed non-coding RNAs induced by transmissible gastroenteritis virus potentially regulate inflammation and NF-κB pathway in porcine intestinal epithelial cell line[J]. BMC Genomics, 2018, 19 (1): 747. doi: 10.1186/s12864-018-5128-5
55	李宇航, 罗仍卓么, 王兴平, 等. NF-κB信号通路调控奶牛乳腺炎的分子作用机制[J]. 畜牧兽医学报, 2021, 52 (10): 2740- 2752. doi: 10.11843/j.issn.0366-6964.2021.010.005
	LI Y H , LUORENG Z M , WANG X P , et al. Molecular regulatory mechanism of NF-κB signaling pathway regulating mastitis in dairy Cows[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52 (10): 2740- 2752. doi: 10.11843/j.issn.0366-6964.2021.010.005
56	MA X L , ZHAO X M , WANG K L , et al. Identification and analysis of long non-coding RNAs that are involved in inflammatory process in response to transmissible gastroenteritis virus infection[J]. BMC Genomics, 2019, 20 (1): 806. doi: 10.1186/s12864-019-6156-5
57	PAN W Z , DU J , ZHANG L Y , et al. The roles of NF-κB in the development of lung injury after one-lung ventilation[J]. Eur Rev Med Pharmacol Sci, 2018, 22 (21): 7414- 7422.
58	LAWRENCE T . The nuclear factor NF-kappaB pathway in inflammation[J]. Cold Spring Harb Perspect Biol, 2009, 1 (6): a001651.

潜在脱靶位点 Potential off-target sites	序列(5′→3′) Sequence
g3 On-target	TACCTCACTCCCAACGCGGATGG
g5 On-target	GGGGGAACTTGCCGACGACCTGG
g3-OT1	aggCTCACTCCCAACGCtGATGG
g3-OT2	TcCCgCAtTCCCAtCGCGGATGG
g3-OT3	TAtCTCcCTtCCAACGCGGgAGG
g3-OT4	cACCTCcCcCCCAAaGCGGACGG
g3-OT5	TACCaCtCTCCCAACaCtGAAGG
g3-OT6	TtCCTCACgCCCAACGCctAGGG
g3-OT7	TACCTggCTCCCAACGaGGtGGG
g3-OT8	TACCTCAggagCAACGCGGATGG
g5-OT1	GtGaGAAaTTGCCGAaGACCAGG
g5-OT2	aGGcGgACTTGCCGAaGACCAGG
g5-OT3	GGGGcAtCcaGCCGACGACCCGG
g5-OT4	GcGGGAACagGCCGgCGACCTGG
g5-OT5	GGGtGAACgTGCCaAaGACCTGG
g5-OT6	GcaGGAACTTGCtGACGgCCAGG
g5-OT7	tGGGGAACTTGCCGgCGAaaAGG
g5-OT8	GGGGGcACTTGCtGACagCCTGG

引物 Primer	引物序列(5′→3′) Primers sequence
g3-OT1	F: AGAGTTTTGAAGGCCTGGTG	R: TGAAGCTGATGGCACCTTTT
g3-OT2	F: CGCAGGGACAGCTTTTAAGA	R: AGACAAATGGACCCTGGAGA
g3-OT3	F: CCAAGGAATCTCAGACTGGC	R: TCAGAATTTGGCAGCCCTTT
g3-OT4	F: ACATTGAGCTTGTCTGGACC	R: GTCCTGAGCTGGTACGAAAG
g3-OT5	F: GTCACAGGGTTACCTCCTCT	R: GACAAAGATGGAGTGGAGGC
g3-OT6	F: AGGTCAAGATGCTGGACTCT	R: AGTCCATGAAAGGGAGTCCA
g3-OT7	F: CGTTTTACACCCATGGTCCA	R: CTTCCAGTGAGAGCAACTGG
g3-OT8	F: GAACAGCAGAGTCAGAGCTC	R: CCCAAAAGCTCTCTCTCGAC
g5-OT1	F: AAGACTTCATGCCTCAGCTG	R: GGTTTGGGGAGAAGTCACTG
g5-OT2	F: CGAAGGAGAAGAACCAGTGG	R: CCTTCATCAAGACCATCCGG
g5-OT3	F: GTGAGAGGATGGCGAACC	R: GATGCTGAGGGTTGAGTGAG
g5-OT4	F: GCAATGTTCTCGGCCAAAC	R: ACCAGAAGCTTCTCTCTCCA
g5-OT5	F: GTTGTCAGACTCCAACCTGG	R: GCACAGGGTGGTCACATTAA
g5-OT6	F: TGTGCTACTACAACGTGCTG	R: CACGCTGAACACGTGGTAG
g5-OT7	F: GCTTTCCTCACCGGATGAAT	R: AGTATCCTTGTCTGGCCAGT
g5-OT8	F: GCCTATCCCTTCCTTCTTGC	R: TCACAGTGAATTGGACTGCC

[1]	REN Lixin, ZHANG Jingyi, XU Shasha, YANG Liu, ZHANG Xingcui, SONG Zhenhui. Effect of ACE2 on Porcine Intestinal Epithelial Cells Infected with Porcine Epidemic Diarrhea Virus in vitro [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1238-1248.
[2]	JIA Hongmin, MA Yonghang, HE Pingli, QIAO Shiyan. Effects of Excessive Lysine on Weaned Piglets and Their Intestinal Epithelial Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(7): 1912-1926.
[3]	LI Meidi, ZHAO Zengjue, LIU Hanqing, FU Jiali, ZHANG Linghua, WU Li. Porcine Cysteine-rich Intestinal Protein 2 Involved in the Gut Immune: Three-dimensional Modeling, Molecular Characterization and Tissue Distribution of mRNA Expression [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3194-3207.
[4]	WANG Yimeng, LIU Xuejiao, WANG Qian, WEI Qing, DOU Caixia, SHANG Zhiyuan, QIAO Jiayun, LI Haihua. Molecular Mechanism of the Injury of IPEC-J2 Caused by Salmonella via NF-κB/β-catenin Signaling Pathway [J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(1): 235-245.
[5]	WANG Zhongqing, LIN Chunfa, ZHONG Wenjie, LUO Yichen, ZHU Zhaorong, LIU Juan. Effect of Zhu Qin Extractive Fluid on the Proliferation and Transcription of Inflammatory Factors in LPS-injured IPEC-J2 Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1500-1508.
[6]	WANG Xiaopeng, XU Kui, WEI Yinghui, ZHANG Xiuling, LIU Shasha, QIU Yiqing, LIU Ying, ZHAO Haiquan, MU Yulian, LI Kui. Establishment of CD13 Gene Knockout IPEC-J2 Cell Lines Mediated by CRISPR/Cas9 System [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(7): 1319-1327.
[7]	PENG Chenglu, ZHANG Yu, DING Xuedong, LI Yu, FENG Shibin, WANG Xichun, LI Jinchun, WU Jinjie. 7S β-conglycinin Triggered Inflammatory Response in IPEC-J2 Cells through NF-κB Signaling Pathway [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(4): 870-878.
[8]	CHEN Xin-yao, ZHANG Jian-long, DONG Xing, CHEN Jing-jie, HUANG Yi-fan, LI Jian. Effects of Hericium Erinaceus Polysaccharide on Antioxidant Ability and ZO-1 Expression in IPEC-J2 Cells under Oxidative Stress [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(9): 1769-1776.
[9]	LENG Yong, SONG Zhen-hui, QING Jia-chao, ZHAI Shao-hua, MAIMAITI-Aizizi. Expression of M Gene and sM Gene of Porcine Transmissible Gastroenteritis Virus by Baculovirus Expression System and VLPs Assmbly of TGEV [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(9): 1432-1437.
[10]	CHEN De-long, ZHU Hong-liang, XU Guang-yong, WANG Ming, JIANG Jin-qi, QIAO Yu, REN Xiao-ming. Study of Lactobacillus Influence the FAK Phosphorylation and Tight Junction Protein Occludin Expression of IPEC-J2 Cells [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2013, 44(2): 283-288.
[11]	ZHANG Kun;HE Qi-gai. Establishment and Clinical Application of a Multiplex Reverse TranscriptionPCR for Detection of Porcine Epidemic Diarrhea Virus, Porcine TransmissibleGastroenteritis Virus and Porcine Group A Rotavirus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(8): 1001-1005.
[12]	. Cloning and Analysis of a 8.5 kb Sequence from the 3′ End of Transmissible Gastroenteritis Virus Genome [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2007, 38(1): 66-71.
[13]	CHENG Jie;LIU Ji-xing;WU Run;YIN Xiang-ping;LI Bao-yu;LAN Xi;WANG Hui. Cloning and Structural Characterization Analysis on the sM ,M and N Genes of Porcine Transmissible Gastroenteritis Virus [J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2005, 36(7): 695-700.

Characteristics Analysis of TGEV Infection Mediated by IPEC-J2 with Knockout of pAPN Gene

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 9

References 58

Related Articles 13

Recommended Articles

Metrics

Comments