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- W2911343437 abstract "The present protocol was aimed to establish a routine transformation procedurevia Agrobaderium tumefaciens for Labisia puml'la var. pumila. The effect ofdifferent factors on T-DNA transfer by measuring transcient expression levels ofa disarmed strain lBA 4404 harbouring the binary vector pBI121 carryingchimeric glucuronidase (GUS) and neomycin phosphotransferase (NP71l) genesexamined. Parameters optimized were light influence, types of wounding, typesof explant, co-cultivation period, bacterial concentration, shaking influence,addition of glucose, pH, temperature, addition of acetosyringone and addition ofsucrose. Improved transformation frequencies were btained with an A.tumefaciens strain carrying kanamycin-type vir genes and when leaves wereinfected with Agrobaderium cells in the early log growth phase. Optimized cocultivationwas performed in the presence of 251-1g/1 of kanamycin. Methodsused have expressed up to 77.62% positive transformants. This result weredone using leaf explants co-cultivated for two days using a batch ofAgrobaderium grown until giving the reading of 0.9 absorbance taken at600nm wavelength. The leaves were poked using sterile needle and co-cultivatedwith Agrobacterium at room temperature with the addition of0.02mg/1 of acetosyringone, 5g/1 of glucose and 15g/1 of sucrose in the pH 7medium. The mixture was left with 24h lighting and 100rpm aggitation.Expression and presence of transgene was assayed by histochemical test andlater confirmed with polymerase chain reaction. Seven percent transgenic plantswere micropropagated and successfully acclimatised. The protocol is yet t beproven t enable it t be reproducible." @default.
- W2911343437 created "2019-02-21" @default.
- W2911343437 creator A5073228611 @default.
- W2911343437 date "2007-01-01" @default.
- W2911343437 modified "2023-09-27" @default.
- W2911343437 title "Genetic transformation of Labisia Pumila using Agrobacterium Tumefaciens" @default.
- W2911343437 hasPublicationYear "2007" @default.
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