Tags

Type your tag names separated by a space and hit enter

[Cloning and expression of ScFv gene against alpha-toxin of Clostridium perfringens type A].
Sheng Wu Gong Cheng Xue Bao. 2001 Sep; 17(5):543-7.SW

Abstract

The VH and VL genes from a hybridoma cell line producing mouse McAb against alpha-toxin of Clostridium perfringens type A were amplified by RT-PCR. The VH and VL genes were connected thought a flexible linker (Gly4Ser)3 and the VH-linker-VL (ScFv) gene was cloned into a vector pGEM-T. The ScFv gene consists of 726 bp encoding 242 amino acid residues. Both VH and VL genes were confirmed as functionally rearranged mouse immunoglobulin variable region. According to kabat classed method, the VH and VL gene segments belong to mouse Ig heavy chain subgroup II (B) and kappa light chain subgroup III respectively. The ScFv gene was amplified inserted the expression vector pHOG21 and transformed into E coli XL1-BLUE. The ScFv protein was highly expressed in recombinant strain XL1-BLUE (pHOG-2E3) and the expression level of the ScFv was about 25% of total bacteria protein by SDS-PAGE. The neutralization assay showed that the expressed ScFv protein could neutralize the phospholipase C activities of alpha-toxin.

Authors+Show Affiliations

College of Life Science, Hebei Normal University, Shijiazhuang 050016, China.No affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

11797218

Citation

Zhao, B H., and C B. Xu. "[Cloning and Expression of ScFv Gene Against Alpha-toxin of Clostridium Perfringens Type A]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 17, no. 5, 2001, pp. 543-7.
Zhao BH, Xu CB. [Cloning and expression of ScFv gene against alpha-toxin of Clostridium perfringens type A]. Sheng Wu Gong Cheng Xue Bao. 2001;17(5):543-7.
Zhao, B. H., & Xu, C. B. (2001). [Cloning and expression of ScFv gene against alpha-toxin of Clostridium perfringens type A]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 17(5), 543-7.
Zhao BH, Xu CB. [Cloning and Expression of ScFv Gene Against Alpha-toxin of Clostridium Perfringens Type A]. Sheng Wu Gong Cheng Xue Bao. 2001;17(5):543-7. PubMed PMID: 11797218.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Cloning and expression of ScFv gene against alpha-toxin of Clostridium perfringens type A]. AU - Zhao,B H, AU - Xu,C B, PY - 2002/1/19/pubmed PY - 2002/2/28/medline PY - 2002/1/19/entrez SP - 543 EP - 7 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 17 IS - 5 N2 - The VH and VL genes from a hybridoma cell line producing mouse McAb against alpha-toxin of Clostridium perfringens type A were amplified by RT-PCR. The VH and VL genes were connected thought a flexible linker (Gly4Ser)3 and the VH-linker-VL (ScFv) gene was cloned into a vector pGEM-T. The ScFv gene consists of 726 bp encoding 242 amino acid residues. Both VH and VL genes were confirmed as functionally rearranged mouse immunoglobulin variable region. According to kabat classed method, the VH and VL gene segments belong to mouse Ig heavy chain subgroup II (B) and kappa light chain subgroup III respectively. The ScFv gene was amplified inserted the expression vector pHOG21 and transformed into E coli XL1-BLUE. The ScFv protein was highly expressed in recombinant strain XL1-BLUE (pHOG-2E3) and the expression level of the ScFv was about 25% of total bacteria protein by SDS-PAGE. The neutralization assay showed that the expressed ScFv protein could neutralize the phospholipase C activities of alpha-toxin. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/11797218/[Cloning_and_expression_of_ScFv_gene_against_alpha_toxin_of_Clostridium_perfringens_type_A]_ DB - PRIME DP - Unbound Medicine ER -