Our Parts

The main goal of our project was to establish the construct which will help us to degrade 7-ketocholesterol consisting in the use of three specific enzymes, but for further applications we submitted them in single modules. This will serve as the basis of a future library for standardized work related to atherosclerosis.

Cholesterol Oxidase

This enzyme was first detected in Chromobacterium sp. We introduced it in a plasmid backbone with chloramphenicol resistance. Its length is of 1871 nucleotides and its codons were optimized in order to use it on E.coli, it already included a stop codon, it was also modified by the addition of a glycosilation site and the peptide signal of human cathepsin.

Oxoacyl Reductase

This enzyme was detected in Rhodococcus jostii . We introduced it in a plasmid backbone with chloramphenicol resistance. Its length is of 1007 nucleotides and its codons were optimized in order to use it on E.coli, it already included a stop codon, it was also modified by the addition of a glycosilation site and the peptide signal of human cathepsin.

7-dehydratase

This enzyme (7-alpha dehydratase) was detected in Rhodococcus jostii . We introduced it in a plasmid backbone with chloramphenicol resistance. Its length is of 602 nucleotides and its codons were optimized in order to use it on E.coli, it already included a stop codon, it was also modified by the addition of a glycosilation site and the peptide signal of human cathepsin.

Neomycin Resistance

This selective marker was gotten from a plasmid for mammalian expression, its length is of 855 nucleotides,and it was isolated from pcDNA3.1(+)/myc-His A.

BGHPA

Stop coding for eucaryotic cells

PCMV

Promoter from Human Cytomegalovirus, this promoter works only on eucaryotic cells.