Primer Design for Cloning of L-arabinose isomerase Gene from Arthrobacter psychrolactophillus into plasmid pET28a(+)

Abstract

A novel putative L-arabinose isomerase (L-AI) called ApL-AI with the accession number WP_110486392.1 was successfully retrieved from Arthrobacter psychrolactophilus B7 genome (Accession: NZ_QJVC01000021.1) through genome mining analysis. This study aimed to obtain the L-AI gene from the Arthrobacter psychrolactophilus B7 genome and clone it into the pET28a(+) plasmid. The primers pair designed in this study successfully amplified the gene using 60 °C of PCR annealing temperature and supported the gene amplicon to insert into the pET28a(+) to form plasmid pET28a(+)-ApLAI. It was proved by the appearance of a 1557-bp amplification band on the gel electrophoresis. The sequencing analysis also revealed that the gene was inserted in the correct direction, with the gene positioned after the promoter and finished with a terminator. Therefore, the plasmid can be used to express the ApL-AI gene to produce the ApL-AI enzyme for downstream analysis and further prospecting.