Engineered single nucleotide polymorphisms in the mosquito MEK docking site alter Plasmodium berghei development in Anopheles gambiae

Table 2 Primer sequences for site-directed mutagenesis

Plasmid constructs	Resulting amino acid mutations	Primer sequence 5′ ➔ 3′
Docking domain mutations
pMEK3	K3M	F: GACGACGACAAGATGAGTATG ATGACAAAAAACAAACTTAA
		R: TTAAGTTTGTTTTTTGTCATCA TACTCATCTTGTCGTCGTC
pMEK4	K6M	F: CAAGATGAGTAAAATGACAATG AACAAACTTAATTTGACGTTG
		R: CAACGTCAAATTAAGTTTGTTCA TTGTCATTTTACTCATCTTG
pMEK5	K3M	F: GACGACGACAAGATGAGTATG ATGACAATG AACAAACTTAATTTGACGTTG
	K6M	R: CAACGTCAAATTAAGTTTGTTCA TTGTCATCA TACTCATCTTGTCGTCGTC
Catalytic domain mutations
pMEK1	S243E	F: GATTGATGA AATGGCCAATTCTTTTGTAGGTACTCGAAG
		R: CTTCGAGTACCTACAAAAGAATTGGCCATTTC ATCAATC
pMEK2	S243E	F: GATTTCGGCGTTTCCGGTCAGTTGATTGATGA AATGGCCAATGA TTTTGTAGGTACTCGAAG
pMEK3
pMEK4	S247D	R: CTTCGAGTACCTACAAAATC ATTGGCCATTTC ATCAATCAACTGACCGGAAACGCCGAAATC
pMEK5

Paired synthetic primers encoding the engineered SNPs (bold) were used to introduce mutations into the MEK-encoding sequence through mutagenic primer-directed replication of both plasmid strands.

ISSN: 1756-3305