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Isolation and Characterization of str Promoter from Bacteria Escherichia coli DH5α using Reporter Gene AmilCP (Acropora millepora)
Mardalisa M.a, Suhandono S.b, Ramdhani M.b
a Department of Marine Science, Faculty of Fisheries and Marine, Riau University, SimpangBaruPanam, PekanBaru, Indonesia
b Research Expertise Group of Genetics and Molecular Biotechnology, School of Life Science and Technology, ITB, Indonesia
[vc_row][vc_column][vc_row_inner][vc_column_inner][vc_separator css=”.vc_custom_1624529070653{padding-top: 30px !important;padding-bottom: 30px !important;}”][/vc_column_inner][/vc_row_inner][vc_row_inner layout=”boxed”][vc_column_inner width=”3/4″ css=”.vc_custom_1624695412187{border-right-width: 1px !important;border-right-color: #dddddd !important;border-right-style: solid !important;border-radius: 1px !important;}”][vc_empty_space][megatron_heading title=”Abstract” size=”size-sm” text_align=”text-left”][vc_column_text]© 2020 IOP Publishing Ltd. All rights reserved.In heat stress conditions, E. coli will produce EF-Tu protein abundantly under the regulation of str promoter. The str promoter from E. coli is known to have sequences that are recognized by two sigma factors (σ70 and σS). The aim of this research was to isolate and characterize str promoter from E. coli DH5a using reporter gene AmilCP. The str promoter from E. coli DH5α (289 bp) had been isolated by Colony PCR using specific primers. Bioinformatics analysis of str promoter showed high conserved areas (100%) for some kinds of E. coli strains. The str promoter was constructed in pSB1C3 vector by modifying OEPC to regulate reporter gene AmilCP. Qualitatively, the ability of str promoter constructs in regulating AmilCP gene was observed as purple-blue colonies of E. coli. Quantitative analysis was performed by measuring cell extract of E. coli at a wavelength of 588 nm after treated at different temperatures (4, 24, 37, 45, 50, 55 and 60 C) for 1 hour. The ability of str promo ter to express reporter gene AmilCP was compared to one powerful promoter which is the lacUV5 promoter. The results show that both promoters are functional promoters and can be induced by temperature (50 C).[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Author keywords” size=”size-sm” text_align=”text-left”][vc_column_text]Acropora,Bioinformatics analysis,Heat stress condition,Isolation and characterization,Measuring cells,Reporter gene,Sigma factor,Specific primers[/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Indexed keywords” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”Funding details” size=”size-sm” text_align=”text-left”][vc_column_text][/vc_column_text][vc_empty_space][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][vc_empty_space][megatron_heading title=”DOI” size=”size-sm” text_align=”text-left”][vc_column_text]https://doi.org/10.1088/1755-1315/430/1/012014[/vc_column_text][/vc_column_inner][vc_column_inner width=”1/4″][vc_column_text]Widget Plumx[/vc_column_text][/vc_column_inner][/vc_row_inner][/vc_column][/vc_row][vc_row][vc_column][vc_separator css=”.vc_custom_1624528584150{padding-top: 25px !important;padding-bottom: 25px !important;}”][/vc_column][/vc_row]