A suspended cell line from Trichoplusia ni embryos was established, and its susceptibility to Autographa californica multiple nuclear polyhedrosis virus （AcMNPV） infection was investigated. This cell line had charac...A suspended cell line from Trichoplusia ni embryos was established, and its susceptibility to Autographa californica multiple nuclear polyhedrosis virus （AcMNPV） infection was investigated. This cell line had characteristics distinct from the BTI-Tn5B 1- 4 cell line （Tn5B 1-4） from T. ni in growth, and showed approximately the same responses to AcMNPV infection, production of occlusion bodies, and levels of recombinant protein expression. No clumps were observed at maximum cell density at late-log phase in shake-flask or T-flask cultures, and thus the cells represent a useful new contribution for baculovirus research. The cells consist of two major morphological types： approximately 70% spindle-shaped cells and 30% round cells. The cell line was highly susceptible to virus infection and produced around 107 AcMNPV occlusion bodies per cell, on average. Production of β-galactosidase and secreted alkaline phosphatase was high with 3.97 ± 0.13 × 10^4 IU/mL and 3.48 ± 0.40IU/mL, respectively. This cell line may be applicable for studies of scale-up production of viruses or baculovirus-insect cell expression. We also believe the new line can be a source for cell clones with higher production of virus and recombinant proteins compared to the parent or other existing cell lines such as Tn5B 1-4.展开更多
Bacteria have long been the favorite expression system for recombinant protein production.However,the flaw of the system is that insoluble and inactive proteins are co-produced due to codon bias,protein folding,phosph...Bacteria have long been the favorite expression system for recombinant protein production.However,the flaw of the system is that insoluble and inactive proteins are co-produced due to codon bias,protein folding,phosphorylation,glycosylation,mRNA stability and promoter strength.Factors are cited and the methods to convert to soluble and active proteins are described,for example a tiglit control of Escherichia coli milieu,refolding from inclusion body and through fusion technology.展开更多
基金Acknowledgments The authors gratefully acknowledge support from the Chinese National Basic Research Program （973） 2003CB114201, Chinese National Science Foundation Project 30771451, and Boyce Thompson Institute, collaborative research Project BTI-QAU 1232007.
文摘A suspended cell line from Trichoplusia ni embryos was established, and its susceptibility to Autographa californica multiple nuclear polyhedrosis virus （AcMNPV） infection was investigated. This cell line had characteristics distinct from the BTI-Tn5B 1- 4 cell line （Tn5B 1-4） from T. ni in growth, and showed approximately the same responses to AcMNPV infection, production of occlusion bodies, and levels of recombinant protein expression. No clumps were observed at maximum cell density at late-log phase in shake-flask or T-flask cultures, and thus the cells represent a useful new contribution for baculovirus research. The cells consist of two major morphological types： approximately 70% spindle-shaped cells and 30% round cells. The cell line was highly susceptible to virus infection and produced around 107 AcMNPV occlusion bodies per cell, on average. Production of β-galactosidase and secreted alkaline phosphatase was high with 3.97 ± 0.13 × 10^4 IU/mL and 3.48 ± 0.40IU/mL, respectively. This cell line may be applicable for studies of scale-up production of viruses or baculovirus-insect cell expression. We also believe the new line can be a source for cell clones with higher production of virus and recombinant proteins compared to the parent or other existing cell lines such as Tn5B 1-4.
基金supported by Queen Saovabha Memorial Institute,The Thai Red Cross Socity
文摘Bacteria have long been the favorite expression system for recombinant protein production.However,the flaw of the system is that insoluble and inactive proteins are co-produced due to codon bias,protein folding,phosphorylation,glycosylation,mRNA stability and promoter strength.Factors are cited and the methods to convert to soluble and active proteins are described,for example a tiglit control of Escherichia coli milieu,refolding from inclusion body and through fusion technology.
