Isolation and sequencing of Folic acid production gene from soil bacilli and its cloning in Escherichia coli

Folic acid or vitamin B9 is one of the most essential vitamins, which plays a key role in the transport of carbon and many biochemical pathways and its deficiency can lead to neurological diseases, anemia and developmental problems in children. The present study aimed to take a step towards producing this vitamin from native strains by isolating and sequencing the folic acid production gene from soil isolated bacillus and cloning it in Escherichia coli.

In this study, 40 samples were collected from 5-10 cm depth from different areas around Sirjan city. Bacillus bacteria were isolated by molecular and biochemical methods. By PCR method, bacteria with FOLA gene were identified and the amplified gene was cloned into E. coli and sequenced. Gene expression was then evaluated by Real time PCR and its phylogenetic relationships were determined.

As a result of screening of 40 soil samples sent to the laboratory, a total of 12 suspected colonies were isolated from Gram-positive bacilli which were identified based on morphological, microscopic and biochemical characteristics and all species were identified as having FOLA gene. FOLA cloning was performed successfully and confirmed its gene expression.

In the present study, the FOLA gene from Bacillus Halotolerance isolated from the soils of the surrounding areas of Sirjan was successfully cloned into PTG19-T expression vector and its successful expression showed that it could be expanded by this method for industrial production of Folate.