انتقال ژن آنتي ژن سطحي ويروس هپاتيت( B (HBsAg به گياه فلفل دلمه اي ﴿‍.Capsicum annum L﴾

چكيده انگليسي :

Transformation of Hepatitis B Virus surface antigen gene into pepper plant Capsicum annuum L Majedeh neisi m neisi@ag iut ac ir Date of Submission Department of Agricaltural Biotechnology Isfahan University of Technology Isfahan Iran Degree M Sc Language Farsi B E Sayed Tabatabaei Prof Supervisor sayedt@cc iut ac ir C Ghobadi Assist Prof Supervisor cyrus@cc iut ac ir Abstract The Hepatitis B virus HBV infection is one of the most viral infections of humans and causes acute and chronic hepatitis and hepatocellular carcinoma The effective way to treatment and prevention of the disease is vaccination Since the chemical drugs are very expensive so in recent years the transgenic plants as an alternative bioreactors for the production of cheap mass and safety and delivery of various recombinant biopharmaceutical compounds and many kind of therapeutic recombinant proteins have been interest to many researchers The HBsAg produced in transgenic plants is antigenically and physically similar to the HBsAg particles derived from human serum and recombinant yeast which are used as vaccines In this study Hepatitis B surface antigen gene HBsAg was transferred in pepper plants that have special significance the area under cultivation nutritional and medicinal value of horticultural products First the sequence of HBsAg gene was idenified by NCBI database For amplification of the target gene by polymerase chain reaction specific primers were designed by oligo analyser software Then to increase the number of pCAMBIA plasmid containing HBsAg gene and easy access to it in emergency condition cloning was done in E coli JM To produce the transgenic plants containing the plasmid containing HBsAg gene transferred to Agrobacterium tumefaciens First for of callus induction and plant regeneration of pepper plant in the success transfer of HBsAg gene to ultra plant tissue culture conditions were optimized in order to select the best explant and hormon mixture Then the target gene was transferred to pepper plant by Agrobacterium mediated The results of the optimization of tissue culture show that mg l BAP mg l IAA for callus induction and mg l BAP mg l IAA are suitable for regeneration Cotyledon explants from week old seedlings are the best selected explants and the appropriate age inoculated with Agrobacterium containing pCAMBIA HBsAg To select transgenic plants the best concentration of selectivity antibiotic Hygromycin was diagnosed mg l In addition use of mg l Zea stimulate the growth of buds were generated from the transformed callus and escalated shooting To induce elongation and inhibition of rosette plants mg l BAP GA were used inculture medium Transfer The regenerated shoots to medium containing coal be caused their rooting after weeks After the suitable growth of pepper plantlets about cm they were transferred to situation in vivo pots PCR reactions showed that there is at least one copy of the HBsAg gene in transgenic pepper plants The results of RT PCR reactions confirmed that the gene is expressed in transgenic pepper These results indicate that the first step to production of edible vaccine against Hepatitis B by produce recombinant proteins in a plant bioreactor of pepper is properly removed Keywords Gene transformation Agrobacterium Hepatitis B surface antigen Transgenic pepper Edible vaccine PDF created with pdfFactory trial version www pdffactory com

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انتقال ژن آنتي ژن سطحي ويروس هپاتيت( B (HBsAg به گياه فلفل دلمه اي ﴿‍.Capsicum annum L﴾

اگروباكترويوم , واكسن خوراكي