Journal of Biosciences

, Volume 32, Issue 4, pp 705–712

Molecular cloning and expression of the C-terminus of spider flagelliform silk protein from Araneus ventricosus

  • Kwang Sik Lee
  • Bo Yeon Kim
  • Yeon Ho Je
  • Soo Dong Woo
  • Hung Dae Sohn
  • Byung Rae Jin
Article

DOI: 10.1007/s12038-007-0070-8

Cite this article as:
Lee, K.S., Kim, B.Y., Je, Y.H. et al. J Biosci (2007) 32: 705. doi:10.1007/s12038-007-0070-8

Abstract

A cDNA coding for the C-terminus of spider flagelliform silk protein (AvFlag) was cloned from Araneus ventricosus. Analysis of the cDNA sequence shows that the C-terminus of AvFlag consists of 167 amino acids of a repetitive region and 87 amino acids of a C-terminal non-repetitive region. The peptide motifs found in spider flagelliform silk proteins, GPGGX and GGX, were conserved in the repetitive region of AvFlag. Phylogenetic analysis further confirmed that AvFlag belongs to the spider flagelliform silk proteins. The AvFlag cDNA was expressed as a 28 kDa polypeptide in baculovirus-infected insect cells. As a new expression approach for spider silk protein, the combination of polyhedrin and AvFlag creates a polyhedrin AvFlag fusion protein (61 kDa) that is produced as recombinant polyhedra; this provides a basis for the source of spider silk proteins for various applications.

Keywords

Araneus ventricosus baculovirus expression vector cDNA cloning flagelliform silk protein spider 

Abbreviations used

BSA

bovine serum albumin

EST

expressed sequence tag

AvFlag

flagelliform silk protein from A. ventricosus

HRP

horseradish peroxidase

MOI

multiplicity of infection

PAUP

Phylogenetic Analysis Using Parsimony

PFU

plaque-forming units

PAGE

polyacrylamide gel electrophoresis

Copyright information

© Indian Academy of Sciences 2007

Authors and Affiliations

  • Kwang Sik Lee
    • 1
  • Bo Yeon Kim
    • 1
  • Yeon Ho Je
    • 2
  • Soo Dong Woo
    • 3
  • Hung Dae Sohn
    • 1
  • Byung Rae Jin
    • 1
  1. 1.College of Natural Resources and Life ScienceDong-A UniversityBusanKorea
  2. 2.School of Agricultural BiotechnologySeoul National UniversitySeoulKorea
  3. 3.Department of Plant MedicineChungbuk National UniversityCheongjuKorea