3381

3054

كارشناسي ارشد

بيوتكنولوژي كشاورزي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1385

دوازده ،114، [II] ص.: مصور،جدول

ص.ع. به فارسي و انگليسي

علي هاتف سلمانيان ، سيروس قبادي

هاله هاشمي سهي

بدرالدين ابراهيم سيد طباطبايي، حميدرضا رحماني

مهندسي كشاورزي

ID3054

به فارسي و انگليسي : قابل رويت در نسخه ديجيتال

127Abstract Plants provide an inexpensive system for the large scale production of valuablerecombinant proteins The ability to carry out post translational modifications similar toother higher eukaryotes and absence of human pathogens endotoxins prions andoncogenic sequences are some of the advantages of plants compared with otherproduction systems In this research first the coding sequence of human growthhormone mature protein substituted gus gene in pBI121 and become under the controlof CaMV35S promoter and nos terminator Agrobacterium tumefaciens GV3101 wastransformed with this construct and be used for transformation of tomato explants Cotyledons and hypocotyls were used as explants and excised from tomato seedlings10 14 days after culture of sterilized seed on MS medium Explants co cultivated withAgrobacterium for two days and then transferred to the selective regeneration mediumcontaining kanamycin Regenerated shoots with appropriate size cultivated in rootingmedium PCR reaction with specific primers from genomic DNA extracted fromregenerated plants and western blot analysis of their proteins confirmed the presenceand expression of human growth hormone gene in transgenic tomato plants The resultsalso illustrated that Zeatin riboside is more effective than Zeatin in tomato regeneration Chloroplast targeting of Growth Hormone protein was the next step of this study Chloroplastic signal sequence of RUBISCO small subunit of rapeseed was used forchloroplast targeting and fused to the sequence of mature human Growth Hormone bySOEing PCR This is an easy and low cost method to attachment of DNA sequences forproduction of fused proteins that can be used in various investigations The fusedfragment substituted the gus gene in pBI121 and used for Agrobacterium mediatedtobacco transformation Western blot analysis shows the expression of the construct intransgenic tobacco plant Correct digestion of signal peptide from junction point toGrowth Hormone by plastid signal peptidase system accumulates mature humanGrowth Hormone without initial Methionine and similar to natural form in human inplant chloroplasts Organelle targeting of recombinant protein cause accumulation convenience of extraction correct folding and posttranslational modification ofproteins It also protects recombinant proteins against cytosolic proteases and increaseproduction in plants Organelle targeting prevents toxic accumulation of recombinantproteins in cytoplasm and protein secretion to the apoplast phloem or xylem wherecontact with environment including microbes and insect pests and minimized negativeeffects by restricting the exposure of environment and non target organisms torecombinant proteins

علي هاتف سلمانيان ، سيروس قبادي

هاله هاشمي سهي

بدرالدين ابراهيم سيد طباطبايي، حميدرضا رحماني