Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons

International Journal of Molecular Sciences
Wenjing ChenQingyou Xia

Abstract

Human platelet derived growth factor (PDGF) is a major therapeutic protein with great demand in the clinical setting; however, its rate of supply is far from meeting needs. Here, we provide an effective strategy to produce PDGF-BB in large quantities using a transgenic silkworm. The codon-optimized PDGF-B gene regulated by the highly efficient sericin-1 expression system was integrated into the genome of a silkworm. The high transcriptional expression of the PDGF-BB gene in the transgenic silkworm competitively inhibited the transcription expression of the endogenous sericin-1 gene which caused a significant 37.5% decline. The PDGF-BB synthesized in the middle silk gland (MSG) of transgenic silkworms could form a homodimer through intermolecular disulfide bonds, which is then secreted into sericin lumen and finally, distributed in the sericin layer of the cocoon. In this study, a protein quantity of approximately 0.33 mg/g was found in the cocoon. Following a purification process, approximately 150.7 μg of recombinant PDGF-BB with a purity of 82% was purified from 1 g of cocoons. Furthermore, the bioactivity assays showed that the purified recombinant PDGF-BB was able to promote the growth, proliferation and migration of NIH/3T...Continue Reading

References

Jan 11, 2001·Development Genes and Evolution·C Horn, E A Wimmer
Jan 24, 2002·Insect Biochemistry and Molecular Biology·J L ThomasG Chavancy
Dec 17, 2002·Nature Biotechnology·Masahiro TomitaKatsutoshi Yoshizato
Sep 4, 2003·Development·Renée V Hoch, Philippe Soriano
Mar 6, 2007·Biochemical and Biophysical Research Communications·H KuriharaK Yamada
Apr 7, 2007·Transgenic Research·Masahiro TomitaKatsutoshi Yoshizato
Jan 27, 2009·Journal of Orthopaedic Research : Official Publication of the Orthopaedic Research Society·Loay Al-ZubeSheldon S Lin
Sep 11, 2009·The FEBS Journal·Masashi IizukaMasahiro Tomita
Dec 25, 2010·Biotechnology Letters·Masahiro Tomita
Jan 16, 2015·Molecular Genetics and Genomics : MGG·Feng WangQingyou Xia
May 5, 2016·Journal of Cosmetic Dermatology·Husein Husein El Hadmed, Rafael Fernandez Castillo
May 11, 2016·Proceedings of the National Academy of Sciences of the United States of America·Jonathan A KlugeDavid L Kaplan
Nov 5, 2016·International Journal of Biological Macromolecules·Lian DuanJiao Li
Feb 13, 2017·Insect Biochemistry and Molecular Biology·Sanyuan MaQingyou Xia
Mar 8, 2017·Gene·Andrius Kazlauskas
Jun 3, 2017·Biomedical Materials·Michaela BienertS Neuss
Jun 8, 2017·Journal of Cellular Physiology·Maryam MajidiniaBahman Yousefi
Jul 20, 2017·Nature Communications·Ulyana ShimanovichTuomas P J Knowles
Feb 7, 2018·Acta Biomaterialia·James Sparkes, Chris Holland

❮ Previous
Next ❯

Citations

Mar 12, 2020·Annual Review of Biomedical Engineering·Jung Woo LeemYoung L Kim
Dec 6, 2019·Acta Biomaterialia·Dimple Chouhan, Biman B Mandal
Jan 16, 2021·International Journal of Molecular Sciences·Tong ZhouZongbin Cui

❮ Previous
Next ❯

Methods Mentioned

BETA
transgenic
PCR
ion affinity chromatography
electrophoresis
fluorescence microscopy

Software Mentioned

BandScan5
Image J

Related Concepts

Related Feeds

Cell Migration

Cell migration is involved in a variety of physiological and pathological processes such as embryonic development, cancer metastasis, blood vessel formation and remoulding, tissue regeneration, immune surveillance and inflammation. Here is the latest research.