Cloning Of Peanut Ara H 1 Gene Promoter And The Research On Gene Family Encoding Storage Proteins By Fiber-FISH

Peanut is an important oil crops. The research on genes related to development of peanut seeds, especially on regulation mechanism of accumulation process of the storage materials establishes a foundation to the application in agricultural production. Peanut is also a source of plant protein for nutrition. The study of the gene structure and expression regulation of peanut storage protein provides an academic base on improving the quality of peanut, breeding peanut variety with high quality and producing hypoallergenic peanuts.The promoter of the gene encoding the major peanut allergy protein Ara h 1 has been cloned with a pair of special primers using Polymerase Chain Reaction technique (PCR). The promoter we have got is 1957 bp. Basically, it is consistent with the sequence published. By replacing the 35S promoter in pBIN 35S-mGFP4 vector with this promoter, we have constructed a plant transgenic expression vector of peanut named pGA1. It contains seed specifically expressed elements, enhancers and some elements regulated by transcription factors. Thus the heterologous gene can be transferred to peanut and expresses in seed easily. The research lays a foundation for establishing the peanut bioreactor.We have also done some initiative researches on the arrangement of gene family encoding peanut storage proteins. We use the technique of fiber fluorescent in situ hybridization (Fiber-FISH) and two promoters of peanut storage protein genes-Arah1 and Arah3 as probes in order to find the DNA region in peanut genome which hybridizes to the two probes. The results indicate that the genes encoding peanut storage protein may exist as some clusters in the peanut chromosomes. The longest continuous fragment contains eleven clusters and covering a 86.78 micrometer region on the DNA fiber corresponding to 217.82 kilo base pairs on the DNA sequence. The average length of the intervals among these eleven clusters is 3.94 micrometer with variance of 0.78 micrometer corresponding to 7.93 kilo base pairs to 11.85 kilo base pairs on the DNA sequence. Based on a large amount of analysises of the dual-colored hybridization signals, we have found that the average length of each cluster was 5.7 micrometer with variance of 1.32 micrometer corresponding to 10.99 kilo base pairs to 17.62 kilo base pairs on the DNA sequence.