Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (5): 92-97.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0902
Previous Articles Next Articles
ZHANG Xi-xi1(), ZHANG Yi-qing2,3, LI Yu-lin3, HAN Xiao3, WANG Guo-qiang3,4, WANG Xiao-jun5, WANG Xu-dong5(), WANG Yun-long2,3()
Received:
2020-07-18
Online:
2021-05-26
Published:
2021-06-11
Contact:
WANG Xu-dong,WANG Yun-long
E-mail:biozxx@126.com;davidwang96@yahoo.com;biowyl@126.com
ZHANG Xi-xi, ZHANG Yi-qing, LI Yu-lin, HAN Xiao, WANG Guo-qiang, WANG Xiao-jun, WANG Xu-dong, WANG Yun-long. Prokaryotic Expression,Purification and Application of N Protein C-terminal Recombinant Protein in Novel Coronavirus(SARS-CoV-2)[J]. Biotechnology Bulletin, 2021, 37(5): 92-97.
引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|
NC-F | GGAATTCCATATGCATCATCACCATCA |
NC-R | CCGCTCGAGTTACGGAAAGGTTTTAT |
Table 1 Primer sequence
引物名称Primer name | 引物序列Primer sequence(5'-3') |
---|---|
NC-F | GGAATTCCATATGCATCATCACCATCA |
NC-R | CCGCTCGAGTTACGGAAAGGTTTTAT |
Fig.2 N protein C-terminal sequence gene expression vector PCR and double enzyme digestion A: M1: DNA molecular weight standard, 1: PCR amplification product. B: M2: DNA molecular weight standard, 2: Double digestion product
Fig.3 Soluble expression of recombinant protein M: The molecular weight standard of protein. 1:Pre-induced sample. 2: Supernatant induced at 37℃. 3: Precipitation induced at 37℃. 4: Supernatant induced at 25℃. 5: Precipitation induced at 25℃. 6: Supernatant induced at 16℃. 7: Precipitation induced at 16℃
Fig.4 SDS-PAGE analysis of prokaryotic expression and purification of the C-terminal recombinant protein of SARS-CoV-2 N protein M: The molecular weight standard of protein. 1:Pre-induced sample.2: Supernatant induced at 37℃. 3:Ni2+ affinity chromatography purification.4: Ni2+ affinity chromatography purification and gel chromatography purification
Fig.6 Western blotting analysis of SARS-CoV-2 N protein C-terminal recombinant protein M: The molecular weight standard of protein. 1: The purified recombinant protein.2: Pre-induced sample
[1] | Liu WB, Liu L, Kou GM, et al. Evaluation of nucleocapsid and spike protein-based enzyme-linked immunosorbent assays for detecting antibodies against SARS-CoV-2[J]. J Clin Microbiol, 2020,58(6):e00461-20. |
[2] |
Sarma P, Shekhar N, Prajapat M, et al. In-silico homology assisted identification of inhibitor of RNA binding against 2019-nCoV N-protein(N terminal domain)[J]. J Biomol Struct Dyn, 2021,39(8):2724-2732.
doi: 10.1080/07391102.2020.1753580 URL |
[3] |
Vasan S, Cui H, Gao Z, et al. Structural genomics of SARS-CoV-2 indicates evolutionary conserved functional regions of viral proteins[J]. Viruses, 2020,12(4):360.
doi: 10.3390/v12040360 URL |
[4] |
Ahmed SF, Quadeer AA, Mckay MR. Preliminary identification of potential vaccine targets for the COVID-19 coronavirus(SARS-CoV-2)based on SARS-CoV immunological studies[J]. Viruses, 2020,12(3):254.
doi: 10.3390/v12030254 URL |
[5] |
Kandeel M, et al. From SARS and MERS CoVs to SARS-CoV-2:moving toward more biased codon usage in viral structural and nonstructural genes[J]. J Med Virol, 2020,92(6):660-666.
doi: 10.1002/jmv.v92.6 URL |
[6] | Yamaoka Y, Jeremiah SS, Miyakawa K, et al. Whole nucleocapsid protein of SARS-CoV-2 may cause false positive results in serological assays[J]. Clin Infect Dis, 2020, 23:ciaa637. |
[7] |
Mu J, Xu J, Zhang L, et al. SARS-CoV-2-encoded nucleocapsid protein acts as a viral suppressor of RNA interference in cells[J]. Sci China Life Sci, 2020,63(9):1413-1416.
doi: 10.1007/s11427-020-1692-1 URL |
[8] | Wen F, Yu H, Guo J, et al. Identification of the hyper-variable genomic hotspot for the novel coronavirus SARS-CoV-2[J]. J Infect, 2020,80(6):671-693. |
[9] |
Gopal GJ, et al. Strategies for the production of recombinant protein in Escherichia coli[J]. Protein J, 2013,32(6):419-425.
doi: 10.1007/s10930-013-9502-5 URL |
[10] | Yin X, Wang W, Tian R, et al. Prokaryotic expression, purification and antigeNi2+city identification of recombinant human survivin protein[J]. Chinese Journal of Cellular & Molecular Immunolog, 2013,29(8):877-881. |
[11] | 王云龙, 米亚双, 等. CA19-9时间分辨荧光免疫层析检测方法的建立[J]. 生物工程学报, 2018,34(6):1012-1018. |
Wang YL, Mi YS. et al. Establishment of a time-resolved fluorescence immunochromatography assay for CA19-9[J] Chinese Journal of Biotechnology. 2018,34(6):1012-1018. | |
[12] | 石英杰, 等. HPV16/18 E6蛋白荧光免疫层析联合检测方法的建立[J]. 免疫学杂志, 2018,34(6):534-538, 546. |
Shi YJ, Wang YL, Ming L, et al. Establishment of a combined method for detection of HPV16/18 E6 protein by fluorescence immunochromatography[J]. Immunological Journal, 2018,34(6):534-538, 546. | |
[13] | Burbelo PD, Riedo FX, et al. Detection of nucleo capsid antibody to SARS-CoV-2 is more sensitive than antibody to spike protein in COVID-19 patients[J]. J Infect Dis, 2020 : 20071423. |
[14] |
Zeng W, Liu G, Ma H, et al. Biochemical characterization of SARS-CoV-2 nucleocapsid protein[J]. Biochem Biophys Res Commun, 2020,527(3):618-623.
doi: 10.1016/j.bbrc.2020.04.136 URL |
[15] |
Tilocca B, Soggiu A, Sanguinetti M, et al. Comparative computational analysis of SARS-CoV-2 nucleocapsid protein epitopes in taxonomically related coronaviruses[J]. Microbes Infect, 2020,22(4-5):188-194.
doi: 10.1016/j.micinf.2020.04.002 URL |
[1] | MEI Huan, LI Yue, LIU Ke-meng, LIU Ji-hua. Study on the Biosynthesis of l-SLR by Efficient Prokaryotic Expression of Berberine Bridge Enzyme [J]. Biotechnology Bulletin, 2023, 39(7): 277-287. |
[2] | LI Wen-shuo, WANG Lin-song, DU Gui-cai, GUO Qun-qun, ZHANG Ting-ting, YANG Hong LI Rong-gui. Gene Cloning of an Aldehyde Dehydrogenase from Bursaphelenchus xylophilus and Biochemical Characterization [J]. Biotechnology Bulletin, 2022, 38(9): 207-214. |
[3] | SUO Qing-qing, WU Nan, YANG Hui, LI Li, WANG Xi-feng. Prokaryotic Expression,Antibody Preparation and Application of Rice Caffeoyl Coenzyme A-O-methyltransferase Gene [J]. Biotechnology Bulletin, 2022, 38(8): 135-141. |
[4] | QIN Xue-jing, WANG Yu-han, CAO Yi-bo, ZHANG Ling-yun. Prokaryotic Expression and Preparation of Polyclonal Antibody of PwHAP5 Gene in Picea wilsonii [J]. Biotechnology Bulletin, 2022, 38(8): 142-149. |
[5] | WANG Guang-li, FAN Chan, WANG Hui, LU Hui-fang, XIA Ling-yin, HUANG Jian, MIN Xun. Prokaryotic Expression,Purification,Identification,and Polyclonal Antibody Preparation of Vibrio cholerae Hemolysin HlyA [J]. Biotechnology Bulletin, 2022, 38(7): 269-277. |
[6] | YI Fang, LAI Peng-cheng, ZHENG Xi-ao, HU Shuai, GAO Yan-li. Research on the Preparation and Purification of Kod DNA Polymerase [J]. Biotechnology Bulletin, 2022, 38(5): 183-190. |
[7] | WANG Jia-li, HE Si-qi, KANG Zi-xi, WANG Jian-xun. Antibody Phage Display Technology and Its Application in the Discovery of Anti-SARS-CoV-2 Antibodies [J]. Biotechnology Bulletin, 2022, 38(5): 248-256. |
[8] | WANG Qiao-ju, HU Yu-meng, WEN Ya-ya, SONG Li, MENG Chuang, PAN Zhi-ming, JIAO Xin-an. Expression and Activity Identification of SARS-CoV-2 S1 Protein [J]. Biotechnology Bulletin, 2022, 38(3): 157-163. |
[9] | LIU Xiao-mei, WANG Dong-xin, ZHANG Chun, WEI Shuang-shi. Inhibition of AAV-mediated RNAi to SARS-CoV-2 S Gene Expression [J]. Biotechnology Bulletin, 2022, 38(3): 188-193. |
[10] | WANG Xiao-qin, HUANG Yin-ping, WANG Wei-qian, WU Ping, QUAN Shu. Expression and Purification of the MLL3SET Protein with a Site-directed Mutation of an Unnatural Amino Acid [J]. Biotechnology Bulletin, 2022, 38(3): 194-202. |
[11] | ZHANG Feng-wen, ZHOU Li-ya, DONG Chao, SHI Yan-mao. Purification of Antioxidant Peptides from Natto Supernatant and Study on Its Activity [J]. Biotechnology Bulletin, 2022, 38(2): 158-165. |
[12] | CHEN Duo, LIU Yong-zhe. Prokaryotic Expression,Purification and Crystallization of N-terminal Domain of Nucleocapsid Protein in SARS-CoV-2 [J]. Biotechnology Bulletin, 2022, 38(12): 149-155. |
[13] | SHEN Jun-qiang, ZHANG Li-ping, YU Rui-ming, WANG Yong-lu, PAN Li, LIU Xia, LIU Xin-sheng. Porcine Kobuvirus Structural Proteins VP0 and VP1 Prokaryotic Expression and Establishment of Indirect ELISA Method [J]. Biotechnology Bulletin, 2022, 38(10): 243-253. |
[14] | CAO Ru-fei, LI Ze-xuan, XU Huan, ZHANG Sha, ZHANG Min-min, DAI Feng, DUAN Xiao-lei. Expression,Purification,and Crystallization of Pif1 Helicase from Bacteroides fragilis [J]. Biotechnology Bulletin, 2021, 37(9): 180-190. |
[15] | TIAN Jia-hui, FENG Jia-li, LU Jun-hua, MAO Lin-jing, HU Zhu-ran, WANG Ying, CHU Jie. Isolation,Purification and Characterization of Laccase LacT-1 from Cerrena unicolor [J]. Biotechnology Bulletin, 2021, 37(8): 186-194. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||