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S -Acyltransferase gene ( pat ) encodes a protein acyltransferase (PAT) that acts as an enzyme catalyzing an acyl chain transfer to the molecular acceptor, mono- or di-acylglycerol, in the biosynthesis of triacylglycerol (TAG). To date,  pat  from tropical marine microalgae has yet to be explored. Here, we identified a marine microalgae isolate (CBI) based on its physical morphology and 18S rRNA gene, then we isolated the  pat  fragment by reverse transcription (RT) PCR, followed by ligation into pGemT vector and cloning in  E. coli  TOP10. The  pat  fragment was characterized using several DNA analysis softwares. Our results showed that the isolate CBI obtained from Maluku waters was green-colored and spherical in shape with a cell diameter of 2–10 μm. A 1644-bp of 18S rRNA gene was obtained and showed a 99% nucleotide sequence similarity with 18S rRNA gene of  Chlorella vulgaris  (GenBank no. KF574391.1). The microalgae isolate was identified as  Chlorella vulgaris  CBI. Reverse transcription produced a 600-bp cDNA, in which the first 300-bp nucleotide was unidentified, however starting from nucleotide no. 301 to the end, it showed a 51% identity with the  pat  fragment of  Chlorella variabilis  (GenBank no. XP_005849764.1). A 77-residue peptide chain deduced was identified as the first N-terminus of PAT (domain transporter protein). This  pat  fragment of  Chlorella vulgaris  CBI was useful as a starting point to isolate and characterize the whole  pat  sequence.

Characterization of S-Acyltransferase Gene Fragment from an Isolate of Tropical Marine Microalgae Chlorella vulgaris CBI