DNA molecules encoding ligand gated ion channels from
The present invention relates in part to isolated nucleic acid molecules (polynucleotides) which encode Dermacentor variabilis ligand gated ion channel proteins. The present invention also relates to recombinant vectors and recombinant hosts which contain a DNA fragment encoding D. variabilis LGIC/GluCl channels, substantially purified forms of associated D. variabilis channel proteins and recombinant membrane fractions comprising these proteins, associated mutant proteins, and methods associated with identifying compounds which modulate associated Dermacentor variabilis LGIC/GluCl, which will be useful as insecticides and acaracides.
1. A purified nucleic acid molecule encoding a D. variabilis LGIC/GluCl channel protein, wherein said nucleic acid molecule comprises a nucleic acid molecule which encodes the amino acid sequence as set forth in SEQ ID NO: 2.
2. A purified nucleic acid molecule encoding a D. variabilis LGIC/GluCl channel protein, wherein said protein comprises the amino acid sequence as set forth in SEQ ID NO:2.
3. An expression vector for expressing a D. variabilis LGIC/GluCl channel protein in a recombinant host cell wherein said expression vector comprises a nucleic acid molecule of claim 2 .
4. A host cell which expresses a recombinant D. variabilis LGIC/GluCl channel protein wherein said host cell contains the expression vector of claim 3 .
5. A process for expressing a D. variabilis LGIC/GluCl channel protein in a recombinant host cell, comprising:
(a) transfecting the expression vector of claim 3 into a suitable host cell; and,
(b) culturing the host cells of step (a) under conditions which allow expression of said D. variabilis LGIC/GluCl channel protein from said expression vector.
6. A purified DNA molecule encoding a D. variabilis LGIC/GluCl channel protein which consists of the nucleotide sequence as set forth in SEQ ID NO:1.
7. The DNA molecule of claim 6 which consists of the nucleotide sequence from nucleotide 170 to nucleotide 1363.
8. A D. variabilis LGIC/GluCl channel protein substantially free from other proteins which comprises the amino acid sequence as set forth in SEQ ID NO:2.
9. A D. variabilis LGIC/GluCl channel protein of claim 8 which is a product of a DNA expression vector contained within a recombinant host cell.
10. A substantially pure membrane preparation comprising the D. variabilis LGIC/GluCl channel protein purified from the recombinant host cell of claim 9 .
11. A D. variabilis LGIC/GluCl channel protein which consists of the amino acid sequence as set forth in SEQ ID NO:2.