Factors Affecting Delivery and Transient Axpression Of Β – Glucuronidase Gene in Denrobium Sonia Ptotocorm-Like-Body.
O. A., Janna, M., Maziah, G. K. A., Ahmad Parveez and K., Saleh (2006) Factors Affecting Delivery and Transient Axpression Of Β – Glucuronidase Gene in Denrobium Sonia Ptotocorm-Like-Body. African Journal of Biotechnology, 5 (2). pp. 88-94. ISSN 1684–5315
The effect ot the biolistic device parameters and other factors affecting delivery and expression of uidA gene in Dendrobium Sonia was investigated. Three week old protocorm like body (PLB) were bombarded with gold microparticles coated with pAHC25 plasmid harbouring the uidA gene which encodes β – glucurinodase. The factors investigated were the helium pressure, target distance, macrocarrier flight distance to stopping screen to target tissue, vacuum pressure, gold mircoparticle size, spermidine and calcium on DNA precipitation, and the number of bombardments. Two days after bombardment, the PLB were subjected to histochemical GUS assay, and transient GUS activity was recorded as blue spots using a Leica stereomicroscope. All the factors tested showed significant effect (p < 0.05) on the delivery of DNA and expression of the uidA gene in Dendrobium PLB except for calcium. Surviving PLBs were able to grow and regenerate normally into plantlets.
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