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Basic Characteristics of Mutations
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Mutation Site
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L469A |
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Mutation Site Sentence
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Coexpressing the mono-site mutant HPV16 L1 L469A with GroEL/ES increased L1-p yield by ~7 fold compared with strains expressing the wild-type L1 gene. |
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Mutation Level
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Amino acid level |
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Mutation Type
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Nonsynonymous substitution |
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Gene/Protein/Region
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L1 |
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Standardized Encoding Gene
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L1
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Genotype/Subtype
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HPV16 |
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Viral Reference
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-
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Functional Impact and Mechanisms
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Disease
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-
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Immune
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- |
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Target Gene
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-
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Clinical and Epidemiological Correlations
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Clinical Information
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- |
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Treatment
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- |
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Location
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- |
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Literature Information
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PMID
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26732286
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Title
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Enhanced expression of soluble human papillomavirus L1 through coexpression of molecular chaperonin in Escherichia coli
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Author
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Pan D,Zha X,Yu X,Wu Y
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Journal
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Protein expression and purification
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Journal Info
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2016 Apr;120:92-8
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Abstract
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The major recombinant capsid protein L1 of human papillomavirus (HPV) is widely used to produce HPV prophylactic vaccines. However, the quality of soluble and active expression of L1 in Escherichia coli was below the required amount. Coexpression with the chaperonin GroEL/ES enhanced L1 expression. Overexpressing GroEL/ES increased the soluble expression level of glutathione S-transferase-fused L1 (GST-L1) by approximately approximately 3 fold. The yield of HPV type 16 L1 pentamer (L1-p) was approximately 2 fold higher than that in a single expression system after purification through size-exclusion chromatograph. The expression and purification conditions were then optimized. The yield of L1-p was enhanced by approximately 5 fold, and those of HPV types 18 and 58 L1-p increased by approximately 3 and approximately 2 folds, respectively, compared with that in the single expression system. Coexpressing the mono-site mutant HPV16 L1 L469A with GroEL/ES increased L1-p yield by approximately 7 fold compared with strains expressing the wild-type L1 gene. L1-p was then characterized using circular dichroism spectra, UV-vis cloud point, dynamic light scattering and transmission electron microscope analyses. Results indicated that the conformation and biological characteristics of L1-p were identical to that of native L1. Hence, overexpressing chaperonin in E. coli can increase the expression level of GST-L1 and L1-p production after purification. This finding may contribute to the development of a platform for prophylactic HPV vaccines.
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Sequence Data
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-
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