Tags

Type your tag names separated by a space and hit enter

[Construction and screening of phage display single chain antibody library against Bursaphelenchus xylophilus cellulase].
Sheng Wu Gong Cheng Xue Bao. 2005 Nov; 21(6):900-5.SW

Abstract

A phage display single-chain variable fragment (scFv) library against Bursaphelenchus xylophilus cellulase (BXC) was constructed and used to screen the specific antibodies binding to BXC. The total RNA was extracted from fresh spleens of BALB/C mice immunized with BXC. Gene fragments encoding VH and VL were amplified by RT-PCR and assembled into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The recombinant fragments were cloned into the phagemids (pCANTABSE) and electroporated into E. coli TG1. The recombinant phagemids were rescued by reinfection of helper phage M13K07. The repertoire of the phage display antibody was about 5 x 10(4). The specific antibodies against BXC were obtained after five rounds of affinity selection. The positive phage clone was used to infect E. coli HB2151. SDS-PAGE and western blot analysis showed that the soluble scFv antibodies expressed bound specifically to BXC. The studies laid foundation for quarantine and pathological study of Bursaphelenchus xylophilu.

Authors+Show Affiliations

College of Life Sciences, Nankai University, Tianjin 300071, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

chi

PubMed ID

16468343

Citation

Tian, Wang, et al. "[Construction and Screening of Phage Display Single Chain Antibody Library Against Bursaphelenchus Xylophilus Cellulase]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 21, no. 6, 2005, pp. 900-5.
Tian W, Zhang Q, Yang WB, et al. [Construction and screening of phage display single chain antibody library against Bursaphelenchus xylophilus cellulase]. Sheng Wu Gong Cheng Xue Bao. 2005;21(6):900-5.
Tian, W., Zhang, Q., Yang, W. B., & Bai, G. (2005). [Construction and screening of phage display single chain antibody library against Bursaphelenchus xylophilus cellulase]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 21(6), 900-5.
Tian W, et al. [Construction and Screening of Phage Display Single Chain Antibody Library Against Bursaphelenchus Xylophilus Cellulase]. Sheng Wu Gong Cheng Xue Bao. 2005;21(6):900-5. PubMed PMID: 16468343.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Construction and screening of phage display single chain antibody library against Bursaphelenchus xylophilus cellulase]. AU - Tian,Wang, AU - Zhang,Qi, AU - Yang,Wen-Bo, AU - Bai,Gang, PY - 2006/2/14/pubmed PY - 2009/7/25/medline PY - 2006/2/14/entrez SP - 900 EP - 5 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 21 IS - 6 N2 - A phage display single-chain variable fragment (scFv) library against Bursaphelenchus xylophilus cellulase (BXC) was constructed and used to screen the specific antibodies binding to BXC. The total RNA was extracted from fresh spleens of BALB/C mice immunized with BXC. Gene fragments encoding VH and VL were amplified by RT-PCR and assembled into a single chain by overlapping PCR with a linker DNA encoding the peptide (Gly4Ser)3. The recombinant fragments were cloned into the phagemids (pCANTABSE) and electroporated into E. coli TG1. The recombinant phagemids were rescued by reinfection of helper phage M13K07. The repertoire of the phage display antibody was about 5 x 10(4). The specific antibodies against BXC were obtained after five rounds of affinity selection. The positive phage clone was used to infect E. coli HB2151. SDS-PAGE and western blot analysis showed that the soluble scFv antibodies expressed bound specifically to BXC. The studies laid foundation for quarantine and pathological study of Bursaphelenchus xylophilu. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/16468343/[Construction_and_screening_of_phage_display_single_chain_antibody_library_against_Bursaphelenchus_xylophilus_cellulase]_ DB - PRIME DP - Unbound Medicine ER -