The Cloning Of Several Genes Which Specific-express In Plant Pod Or Seed-coat

In this study, we cloned three genes that specific-expresse in pod or in seed-coat. The first gene is a seed-coat expressed sequence tag (PCS11) in peanut (Arachis hypogaea L.) , which only expressed in peanut' seed-coat. The second one is a hydrophobic protein gene(HPS) in soybean (Glycine max L.) ,which specific-express in pod. And the third gene is MYB61 that specific-expressed in the Arabidopsis seed-coat. The idiographic experimental results were as follows:1. Cloning of the peanut seed-coat specific expressed sequence tag (PSC11): According to the published sequence of PSC11, we designed a pair of primers to PCR the peanut genomic DNA. The length of PCR's result is about 500bp.we found it has a nucleotide identity of about 93% to the published sequence. By BLAST, we found the sequence has significant similarity to the subtilisin-like protease genes in Arabidopsis thaliana and soybean. The sequence similaritiy suggested that peanut PSC11 was also a subtilisin-like protease. Then the genomic DNA was digested by HinaT.II, then the digestion run through electronphoresis gel, then the DNA fragments were transferred to a NC membrane via upward capillary transferring from Agrose gel. After immobilizing, we used PSC11 as a probe, which labeled with Dig, to hybridize with the DNA in the membrane. From the result, we found the hybridization's signal is a good single band.2.Cloning of Hydrophobic protein gene ( HPS ) in soybean: According to the published sequence of HPS, we designed a pair of primers to amplify soybean's genomic DNA. The result of PCR is about 700bp. we found it has a sequence identity of about 99% to the published sequence. And the amino acid sequence identity is 100% to the published one.4.Molecular cloning of MYB61 gene in Arabidopsis: We designed a pair of primers to PCR the Arabidopsis genomic DNA. The result of PCR is 1357bp in length and the DNA sequence contains two introns.