EurekaMag.com logo
+ Translate

Expression of the vesicular stomatitis virus nucleocapsid protein controlled by the lambda p r and p l promoters in escherichia coli


, : Expression of the vesicular stomatitis virus nucleocapsid protein controlled by the lambda p r and p l promoters in escherichia coli. Journal of Biotechnology 2(5): 289-302

Expression vectors were constructed in which a complementary DNA specifying the vesicular stomatitis virus nucleocapsid (VSV N) protein was inserted near the translational initiation region downstream from the thermoinducible PR or PL promoter of bacteriophage .lambda. Expression of the VSV N-protein was determined by a radioimmunoassay with monoclonal antibody prepared against the VSV N-protein. The expression of the VSV N-protein in E. coli was low with either system. The deletion of a part of leader sequence from the translational initiation signal to the VSV N-gene resulted in at least 30-fold increase in production of the VSV N-protein. The VSV N-protein in E. coli was also analyzed by radioimmune blot after separation of proteins by gel electrophoresis. Degraded proteins reacted with the antibody were also observed in the cell extracts.

(PDF 0-2 workdays service)

Accession: 005432273

Submit PDF Full Text: Here


Submit PDF Full Text

No spam - Every submission is manually reviewed

Due to poor quality, we do not accept files from Researchgate

Submitted PDF Full Texts will always be free for everyone
(We only charge for PDFs that we need to acquire)

Select a PDF file:
Close
Close

Related references

Das, T.; Banerjee, A.K., 1993: Expression of the vesicular stomatitis virus nucleocapsid protein gene in Escherichia coli: analysis of its biological activity in vitro. The nucleocapsid (N) protein genes of vesicular stomatitis virus, serotypes New Jersey and Indiana, were expressed in Escherichia coli BL21(DE3) transformed by the recombinant expression vector pET-3a containing N genes under the control of the T7...

Sprague, J.; Condra, J.H.; Arnheiter, H.; Lazzarini, R.A., 1983: Expression of a recombinant DNA gene coding for the vesicular stomatitis virus nucleocapsid protein. A cDNA clone containing the entire vesicular stomatitis virus nucleocapsid gene was assembled by fusing portions of two partial clones. When the cDNA clone was inserted into a new general-purpose eucaryotic expression vector and introduced into ap...

Rose, J.K.; Shafferman, A., 1981: Conditional expression of the vesicular stomatitis virus glycoprotein gene in Escherichia coli. Bacterial plasmids that directed expression of the vesicular stomatitis virus glycoprotein (G-protein) gene under control of the tryptophan operon regulatory region were constructed. A plasmid directing the synthesis of a G-protein-like protein (c...

Pahari, S.; Pari, A.; Chattopadhyay, D.J., 1996: Periplasmic expression of biologically active vesicular stomatitis virus phosphoprotein P in Escherichia coli. Overexpression of a clone of vesicular stomatitis virus phosphoprotein P (New Jersey serotype) using T7 promoter with phoA leader sequence and a simpler two-step purification procedure of the expressed protein has been developed. The purified prot...

Q.H.iCai; Gong ZhenHua; Yang ZengQi; L.W.i; Liu JunHui; Pan KangSuo; Wang RuoCong; Jiang ZhengJun; Zheng ZengRen, 2005: Highly efficient expression and purification of truncated glycoprotein gene of vesicular stomatitis virus in Escherichia coli. The glycoprotein gene of vesicular stomatitis virus Indiana (VSV-Ind) without signal and transmembrane region was amplified by PCR. The DNA sequenced EcoRI and XhoI restricted core fragment was cloned in a frame downstream of T7 promoter in the pE...

Arnheiter, H.; Davis, N.L.; Wertz, G.; Schubert, M.; Lazzarini, R.A., 1985: Role of the nucleocapsid protein in regulating vesicular stomatitis virus RNA synthesis. We describe experiments with two monoclonal antibodies to the vesicular stomatitis virus (VSV) nucleocapsid protein N with strikingly different characteristics. Antibody 1 binds to nucleocapsids and probably the pool of free (unbound) N protein; i...

Scofield, R.H.; Harley, J.B., 1991: Autoantigenicity of Ro/SSA antigen is related to a nucleocapsid protein of vesicular stomatitis virus. The fine specificity of a reference human anti-Ro/SSA autoantibody-containing serum has been analyzed by using sequential overlapping octapeptides from the human 60-kDa Ro/SSA antigen. From preliminary data, the most antigenic octapeptide in the c...

Bilsel, P.A.; Rowe, J.E.; Fitch, W.M.; Nichol, S.T., 1990: Phosphoprotein and nucleocapsid protein evolution of vesicular stomatitis virus New Jersey. The entire phosphoprotein (P) and nucleocapsid (N) protein gene sequences and deduced amino acid sequences for 18 selected vesicular stomatitis virus isolates representative of the natural genetic diversity within the New Jersey serotype are repor...

Gupta, A.K.; Banerjee, A.K., 1997: Expression and purification of vesicular stomatitis virus N-P complex from Escherichia coli: role in genome RNA transcription and replication in vitro. The nucleocapsid protein (N) and phosphoprotein (P) genes of vesicular stomatitis virus (VSV), Indiana serotype, were coexpressed in Escherichia coli BL21(DE3) by using the expression vector pET-3a. The coexpression resulted in the formation of N-...

Capone, J.; Ghosh, H.P., 1984: Association of the nucleocapsid protein N of vesicular stomatitis virus with phospholipid vesicles containing the matrix protein M. The matrix protein M and the nucleocapsid protein N were isolated from vesicular stomatitis virus and reconstituted into artificial phospholipid vesicles. While the M protein could be reconstituted into phospholipid vesicles, the N protein had no...