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Studies On Hevea Brasiliensis Transformation With Human Serum Albumin CDNA

Posted on:2006-04-13Degree:MasterType:Thesis
Country:ChinaCandidate:M CongFull Text:PDF
GTID:2133360152996397Subject:Biochemistry and Molecular Biology
Abstract/Summary:PDF Full Text Request
Human serum albumin (HSA) is a kind of protein abundant in human blood plasma which can keep osmotic pressure of blood and transfer nutrient from one part of the body to another. It is widely used in clinic. But now most HSA comes from blood plasma. And HSA is restricted in clinic because of shortage of blood source and the latent viruses. Then it is urgent to produce HSA by other means and recombinant HSA from genetic engineering is prosperous now.Compared with microbe bioreactor and animal bioreactor, plant bioreactor has many unique advantages. Hevea brasiliensis has strong laticifer system which is like galacto-phore in animal. Latex runs out after laticifer is cut so that it is easy for proteins contained in latex to be cured and isolated. When proper promoter is added before foreign gene, it can be expressed in laticifer system. Then laticifer system can be turned into an ideal bioreactor. Herein lies the suppose that to establish regeneration system of Hevea brasiliensis and to transfer the expression vector combining REF promoter with HSA cDNA by bombardment or agrobacteria infection. In the study, quite a few of results are got as below:(1) Immature androecia, seeds and cultivated plantlets were used as explants. They were cultured on modified MS medium with different combination of auxin and cytokinin. Then large quantities of calli came out.For immature androecia, the best callus induction medium was MS medium added to 7.0% maltose, 1.2mg/L PVP, 4.0mg/L 2,4-D and 0.2mg/L 6-BA with 1.0% agar-agar powder. The pH of the medium was 5.8. Culture the explants on medium at 25 ± 1 °C in darkness. Carbon source was changed with 7.0% sucrose and the plant hormones were turned down to 1/10 after the first culture without any change of other things.(2)For immature seed, the best induction medium was MS medium with 4.0mg/L 2,4-D,0.2mg/L KT ,1.2mg/L PVP,7.0% sucrose and 1.0% agar-agar powder with pH value of 5.8. Plant hormones were turned down to 1/10 after the first culture.(3)For cultivated plantlet, the best callus induction medium was MS medium with...
Keywords/Search Tags:Hevea brasiliensis, human serum albumin, transformation, callus, embryo, particle gun, agrobacterium
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