4080

3858

كارشناسي ارشد

بيوتكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1386

سيزده، 102،[II] ص: مصور، جدول، نمودار

ص.ع.به: فارسي و انگليسي

رضا زارعي

14/5/87

بدرالدين ابراهيم سيد طباطبايي، نعمت الله اعتمادي

مهندسي كشاورزي

ID3858

به فارسي و انگليسي: قابل رويت در نسخه ديجيتال

AbstractObtaining optimum sensitivity in detection of low abundant proteins is one of majorobstacles in proteomics studies One solution to this problem is to use the naturalfractions present in the cells and indeed nowadays analysis of organelle proteomicsconstitutes a major avenue in this field In this context chloroplast is one the mostimportant plant organelles that has been receiving attention for many reasons Inaddition to supporting photosynthesis chloroplasts also synthesize hormones fattyacids and lipids amino acids vitamins B1 K1 and E nucleotides secondarymetabolites such as alkaloids and isoprenoids and are required for nitrogen and sulfurassimilation Pomegranate Punica granatum that has been under study in this projectis a plant rich of valuable secondary metabolites with significant medical effects such asanticancer antioxidant and antimicrobial activities With remark to the potential valueof pomegranate in producing these compounds in the present study pomegranatechloroplast was isolated by mechanical homogenization followed by Percoll gradient Inaddition tobacco chloroplast was also similarly purified to serve as a model system forisolation and examination of the chloroplasts Both isolated chloroplasts were shown tobe free of other organelles by Western analysis utilizing antibodies specific tomitochondrion cytosol and plasma membrane Comparison of chloroplast isolation inpomegranate and tobacco indicated the presence of interfering phenolic compounds inpomegranate extract that were subsequently dealt with by addition of 5 PVP to thechloroplast isolation buffer It was also found that pomegranate leaves might yield halfthe chloroplast amount compared to that from the tobacco leaves Furthermorepreliminary analysis indicated that the protein profile of pomegranate and tobacco wererather different as judged by SDS PAGE Following pomegranate chloroplast isolationand protein extraction chloroplast proteome was resolved using 2D PAGE from which4 protein spots were isolated and analyzed by de novo sequencing These proteins wereidentified as a 53 kDa ATP synthase beta subunit a probable 70 kDa malatedehydrogenase and a probable 55 kDa Beta amylase Fourth protein spot could not beidentified Literature review shows that this report constitutes the first isolation andproteomics analysis of pomegranate chloroplast

رضا زارعي

بدرالدين ابراهيم سيد طباطبايي، نعمت الله اعتمادي