Difference between revisions of "Part:BBa K4879009"

Latest revision as of 16:16, 11 October 2023

Yarrowia lipolytica HygR construct

The transcriptional unit for HygR, designed for expression in Y. lipolytica in order to delete the faa1 gene from our chassis.

Characterization

Cloning: The construct, along with homologous flanking sequences corresponding to the faa1 gene locus, were synthesized by iGEM’s DNA synthesis partner IDT. The resulting gene fragment was then PCR amplified with the respective primers to attain a higher copy number of the fragment. The amplification of the fragment was verified by running a 0.8% agarose gel, with the fragments expected to show at around the 3000 bp region.

Fig: Lane 1 is the DNA ladder, and lanes 4-7 contain the successfully amplified fragments

The amplified fragment was then gel-extracted and subsequently transformed into our chassis.

Validation: The transformed yeast culture was then plated onto the selection plates containing about 200 µg/ml of hygromycin B and were incubated at 30°C. The growth of the yeast on the selection plates confirms the functional validity of the expression construct.

Fig 2: The growth of transformed Y. lipolytica on the hygromycin B selection plate, confirming part functionality.

Sequence and Features