Partial Sequence of Acid Phosphatase-1^1 Gene (Aps-1^1) Linked to Nematode Resistance Gene (Mi) of Tomato

概要

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A partial amino acid sequence of acid phosphatase-1^1 (apase-1^1), one of acid phosphatase isozymes of tomato, was identified. This information enabled us to synthesize degenerated primer pools of oligonucleotides for polymerase chain reactions (PCR) using cDNA for poly(A)^+ RNA of tomato leaves as a template. As a result, a 135-bp, then a 467-hp PCR product were obtained. Nucleotide sequencing of these two PCR products gave a total of 522-bp sequence that was identified as a part of the Asp-1^1 gene judging from the amino acid sequence deduced from it. Using the 135-bp PCR product as a probe, we detected the restriction fragment length polymorphism (RFLP) in two different lines of tomato by genomic Southern blot analysis. We also did pulsed-field gel electrophoresis (PFGE) and Southern blot analysis to search for suitable fragments to clone into a YAC vector. As a result, a single band with a size that could be cloned into a YAC vector was detected when the genomic DNA was digested with some kinds of restriction enzymes.
社団法人日本農芸化学会の論文
1992-04-23