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Stable Expression and Efficient Secretion of hSCF and hINFγ Protein using Binary Vectors in Chlorella vulgaris

클로렐라에서 바이너리 벡터를 이용한 hSCF와 hINFγ 단백질의 안정적인 발현과 효율적인 분비

  • 정유정 ((주)바이오이즈 중앙연구소) ;
  • 민희경 ((주)바이오이즈 중앙연구소) ;
  • 이원영 ((주)바이오이즈 중앙연구소) ;
  • 김성천 ((주)바이오이즈 중앙연구소)
  • Received : 2024.03.13
  • Accepted : 2024.04.04
  • Published : 2024.06.30

Abstract

Microalgae have great potential in the biomedical and pharmaceutical industries as a new type of bioreactor that can produce proteins for specific purposes, including recombinant proteins, pharmaceuticals, and industrial enzymes. Despite the production advantages and importance of microalgae-based expression systems, studies on secretion efficiency are limited. In this study, for stable expression and efficient secretion of the heterologous protein (human SCF and human INFγ) in Chlorella vulgaris, we constructed SP:hSCF:His and SP:hINFγ:His plant binary vectors using the signal peptide (SP) of Chlamydomonas reinhardtii, and we obtained stable transformants through the effective agrobacterium-mediated transformation of these vectors. Transformants with accurately inserted hSCF and hINFγ demonstrated stably increased mRNA and protein expression using RT-PCR and western blotting under the same culture conditions. Following the analysis of the proteins secreted into the culture medium using ELISA, it was confirmed that hINFγ was effectively produced in the transformed C. vulgaris culture medium. The overall findings indicate that the combination of heterologous protein and SP may be crucial for ensuring the expression and secretion of recombinant proteins in Chlorella culture systems.

Keywords

References

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