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Diversity of G proteins in lepidopteran cell lines: Partial sequences of six G protein alpha subunits

Archives of Insect Biochemistry and Physiology, ISSN: 0739-4462, Vol: 57, Issue: 3, Page: 142-150
2004
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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-like and one Gα-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-like and Gα-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. © 2004 Wiley-Liss, Inc.

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