Cloning the cDNA encoding the AmbtV allergen from giant ragweed (Ambrosia trifida) pollen

Ghosh, Balaram ; Perry, Michael P. ; Marsh, David G. (1991) Cloning the cDNA encoding the AmbtV allergen from giant ragweed (Ambrosia trifida) pollen Gene, 101 (2). pp. 231-238. ISSN 0378-1119

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Ragweed (Ambrosia) pollens contain a number of proteins that cause allergic disease in ragweed-sensitive people. The cloning of the AmbtV cDNA is important, since the 4.4-kDa AmbtV, one of the allergens in giant ragweed (Ambrosia trifida) pollen, serves as a simple model system to study the basic structural requirements for immune recognition of foreign protein allergens. We report the cloning of the AmbtV cDNA by means of the polymerase chain reaction (PCR) using degenerate primers. We generated three sets of overlapping cDNA clones by a combination of PCR and anchored-PCR, and determined the complete nucleotide (nt) sequence. From the nt sequence, the amino acid (aa) sequence of the protein was confirmed and the leader sequence was deduced. This general approach can be used to clone allergen and other cDNAs from complex biological sources provided partial aa sequence information is available. It may be the best available approach in cases where the isolation of clones from a cDNA library is difficult, which proved to be the case for AmbtV.

Item Type:Article
Source:Copyright of this article belongs to Elsevier Science.
Keywords:Recombinant DNA; Polymerase Chain Reaction; Degenerate Oligodeoxyribonucleotide; M13 Vector; cDNA Sequence; Immune Recognition; Allergic Response
ID Code:66080
Deposited On:21 Oct 2011 03:28
Last Modified:21 Oct 2011 03:28

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