New York Structural Biology Center

Jonah Cheung

A method of creating crystals of insect acetylcholinesterase. A polynucleotide is obtained that encodes for acetylcholinesterase in a targeted insect. The polynucleotide contains a catalytic core sequence. A recombinant DNA construct is formed by adding a fusion protein and a polyhistidine tag to the catalytic core sequence. The recombinant DNA construct can be further modified by adding known mutations for resistance to insecticides. A growth medium is transfected with the recombinant DNA construct. A polypeptide encoded by the recombinant DNA construct is separated from the growth medium to form a concentrate. The polyhistidine tag is removed from the concentrate. The concentrate is exchanged into a buffer to create a buffered concentrate. Crystals, suitable for X-ray crystallography are then grown with the buffered concentrate.

System and method for creating crystals of insect acetylcholinesterase

A system and method for creating crystals from human acetylcholinesterase. The crystals can then be analyzed using X-ray crystallography. A segment of DNA for human acetylcholinesterase is isolated that includes the codon sequence GAGGGC. A polyhistidine tag is inserted between codon GAG and codon GGC to produce a recombinant acetylcholinesterase construct. The recombinant acetylcholinesterase construct is used to transfect cells on a prepared growth medium. The growing cells secrete a portion of the recombinant acetylcholinesterase construct. The secreted portion is concentrated and the tag cleaved. The concentrate is then buffered and used to form crystals.

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Brooklyn, NY, United States of America

Jonah Cheung