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Research in Pharmaceutical Sciences، جلد ۱۲، شماره ۲، صفحات ۱۶۸-۱۷۵
عنوان فارسی
چکیده فارسی مقاله
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عنوان انگلیسی High level expression of recombinant human growth hormone in Escherichia coli: crucial role of translation initiation region
چکیده انگلیسی مقاله For high-throughput production of recombinant protein in Escherichia coli ( E. coli ), besides important parameters such as efficient vector with strong promoter and compatible host, other important issues including codon usage, rare codons, and GC content specially at N-terminal region should be considered. In the current study, the effect of decreasing the percentage of GC nucleotides and optimizing codon usage at N-terminal region of human growth hormone (hGH) cDNA on the level of its expression in E. coli were investigated. Mutation in cDNA of hGH was performed through site-directed mutagenesis using PCR. Then, the mutant genes were amplified and cloned into the expression vector, pET-28a. The new constructs were transformed into the BL21(DE3) strain of E. coli and chemically induced for hGH expression. At the final stage, expressed proteins were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS - PAGE), scanning gel densitometry, and western blot. SDS-PAGE scanning gel densitometry assay and western blot analysis revealed higher expression level of hGH by using the two new expressions constructs (mutant genes vectors with decreasing GC content and optimized-codon usage at N-terminal of cDNA) in comparison with wild gene expression vector. Obtained results demonstrated that decreasing the GC nucleotide content and optimization of codon usage at N-terminal of the hGH cDNA could significantly enhance the expression of the target protein in E. coli . Our results highlight the important role of both 5´ region of the heterologous genes in terms of codon usage and also GC content on non-host protein expression in E. coli.
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نویسندگان مقاله مهسا قویم | mahsa ghavim
1damghan branch, islamic azad university, damghan, i.r. iran.

سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)

خلیل abnous | khalil abnous
2pharmaceutical research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

فاطمه آراسته | fatemeh arasteh
2pharmaceutical research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran. 3department of pharmaceutical biotechnology, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

سحر تقوی | sahar taghavi
2pharmaceutical research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

مریم سادات نبوی نیا | maryam sadat nabavinia
3department of pharmaceutical biotechnology, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

مونا علی بلندی | mona alibolandi
2pharmaceutical research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

محمد رمضانی | mohammad ramezani
2pharmaceutical research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran. 3department of pharmaceutical biotechnology, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran. 4nanotechnology research center, school of pharmacy, mashhad university of medical sciences, mashhad, i.r. iran.

سازمان اصلی تایید شده: دانشگاه علوم پزشکی مشهد (Mashhad university of medical sciences)

نشانی اینترنتی http://rps.mui.ac.ir/index.php/jrps/article/view/1757
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/115/article-115-350607.pdf
کد مقاله (doi)
زبان مقاله منتشر شده en
موضوعات مقاله منتشر شده
نوع مقاله منتشر شده Original Article
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