Open AccessCloning Reference Gene rpoB in Serratia plymuthica UBCF_13
Author(s) -
Robi Trivano,
R Fatiah,
J Jamsari
Publication year - 2021
Publication title -
iop conference series. earth and environmental science
Language(s) - English
Resource type - Journals
eISSN - 1755-1307
pISSN - 1755-1315
DOI - 10.1088/1755-1315/741/1/012040
Subject(s) - rpob , gene , housekeeping gene , plasmid , cloning (programming) , clone (java method) , biology , microbiology and biotechnology , genetics , gene expression , computer science , 16s ribosomal rna , programming language
The rpoB gene is one of the housekeeping genes in the Serratia plymuthica UBCF_13. This gene is usually used as a reference gene in qPCR analysis to quantify the expression level of the target gene. In the absolute analysis, the reference gene qPCR was cloned to create a standard curve for calculating the copy number of the target gene expression level. The method used in this study was to clone a partial rpoB gene through the ligation to pGEM®-T Easy plasmid cloning vector and transform it to E. coli Dh5a to proliferation recombinant plasmid. The results of this study indicated the reference rpoB gene was successfully cloned using pGEM®-T Easy with the size of insert gene 118 bp length.