Cloning of α-Amylase enzyme producing gene
into nonα-amylase producing organism and its
Expression

Parvati Saiyam; Kuldeep Diwedi; Sujeet K. Singh; Shruti Shukla; Madhulika Singh

doi:https://doi.org/10.14445/22490183/IJBTT-V4P601

Research Article | Open Access | Download PDF

Volume 4 | Issue 1 | Year 2014 | Article Id. IJBTT-V4P601 | DOI : https://doi.org/10.14445/22490183/IJBTT-V4P601

Cloning of α-Amylase enzyme producing gene into nonα-amylase producing organism and its Expression

Parvati Saiyam , Kuldeep Diwedi , Sujeet K. Singh , Shruti Shukla , Madhulika Singh

Citation :

Parvati Saiyam , Kuldeep Diwedi , Sujeet K. Singh , Shruti Shukla , Madhulika Singh, "Cloning of α-Amylase enzyme producing gene into nonα-amylase producing organism and its Expression," International Journal of Computer Trends and Technology (IJCTT), vol. 4, no. 1, pp. 1-3, 2014. Crossref, https://doi.org/10.14445/22490183/IJBTT-V4P601

Abstract

In this work, the gene of ?-amylase from B. subtilis was cloned and expressed in Non-amylase produsing E.coli. B. subtilis was isolated by performing various biochemical tests. The organism producing ?-amylase enzyme was successfully isolated. The isolation of genomic DNA followed by amplification of Gene Of Interest using ?-amylase primer and annealing temperature 56°C was done using PCR technique. The amplified product was eluted from electrophoresis gel and subjected to restriction digestion (EcoRI) and then ligation with the vector (Plasmid DNA pUC18). A competent cell was prepared using chemical method (CaCl2). The ligation mixture was allowed to react with the competent cells in order to produce transformed cells producing ?-amylase enzyme. The mixture was spread over the starch plates. The successfully transformed E.coli cells were observed over the Starch agar plate.

Keywords

Bacillus subtilis, E.coli, PCR, pUC18, EcoRI, etc.

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