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Largest synthetic gene ever built offers insights into anti-malarial drug resistance

ion with investigators at the National Institutes of Health, described in 2000 that the PfCRT gene is primarily responsible for the vast majority of drug resistance that leads to human death from malaria. Earlier this year, they also helped describe how the gene PfNHE likely contributes, and this turns out not to be as vital as PfCRT. Now, based in part on this study, they know that PfMDR1 is dependent on mutated PfCRT, and probably doesn't act independently.

"The contribution of PfMDR1 to anti-malarial drug resistance is important, but much smaller than what many researchers thought it would be," said Roepe. "Before the discovery of PfCRT, many thought PfMDR1 would be the primary culprit."

Roepe, and his two other co-authors, Georgetown researchers Linda Amoah, PhD, and Jacqueline Lekostaj, an MD/PhD student at Lombardi Comprehensive Cancer Center, employed an unusual method to construct the PfMDR1 gene. They first obtained the PfMDR1 protein sequence from the NIH PubMed data base. They then "back translated" the protein sequence using a computer program to change the parasite gene sequence − which combines nucleotide base pairs in a way that is found in no other life form − into one that could be "read" by yeast. The yeast are then able to produce the PfMDR1 protein, in its proper sequence, from this synthetic gene. They previously used this same technique to create and express a synthetic version of the much smaller PfCRT gene. PfMDR1 is made up of about 4,000 base pairs and its protein is particularly large, Roepe said, making the recent work much more challenging.

Already, analysis of these three anti-malarial drug resistance genes is painting a picture of how the parasite escapes destruction by most of the agents now used against it, he said.,

When a malaria parasite is transmitted through a mosquito bite, the mosquito injects the parasite into skin or blood, which then travels to the liver within minutes, grows
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Source:Georgetown University Medical Center


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