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Diversity of G proteins in Lepidopteran cell lines: Partial sequences of six G protein alpha subunits
Author(s) -
Knight Peter J.K.,
Grigliatti Thomas A.
Publication year - 2004
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.20018
Subject(s) - sf9 , biology , g protein , protein subunit , g alpha subunit , spodoptera , microbiology and biotechnology , biochemistry , receptor , gene , recombinant dna
The aim of this work was to sample the diversity of G protein α subunits in lepidopteran insect cell lines. Here we report the amplification by degenerate PCR of partial sequences representing six G protein α subunits from three different lepidopteran insect cell lines. Sequence comparisons with known G protein α subunits indicate that the Sf9, Ld and High Five™ cell lines each contain (at least) one Gα q ‐like and one Gα i ‐like Gα subunit. All six PCR products are unique at the nucleotide level, but the translation products of the three Gα q‐like partial clones (Sf9‐Gα 1, Ld‐Gα 1, and Hi5‐Gα 1) are identical, as are the translation products of the three Gα i‐like partial clones (Sf9‐Gα 2, Ld‐Gα 2, and Hi5‐Gα 2). Both the Gα q ‐like and Gα i ‐like translation products are identical to known Gα subunits from other Lepidoptera, are highly similar (88–98%) to Gα subunits from other invertebrates including mosquitoes, fruit flies, lobsters, crabs, and snails, and are also highly similar (88–90%) to known mammalian Gα subunits. Identification of G protein α subunits in lepidopteran cell lines will assist in host cell line selection when insect cell lines are used for the pharmacological analysis of human GPCRs. Arch. Insect Biochem. Physiol. 57:142–150, 2004. © 2004 Wiley‐Liss, Inc.