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Title: Standardization of different parameters for ‘particle gun’ mediated genetic transformation of sugarcane (Saccharum officinarum L.)
Authors: Kaur, Ajinder
Gill, M S
Gill, Raman
Gosal, S S
Keywords: embryogenic calli
GUS assay
‘particle gun’ bombardment
Issue Date: Jan-2007
Publisher: CSIR
IPC CodeInt. Cl.⁸ A01H5/00; C12N15/9
Abstract: Three different parameters, viz. quantity of DNA, size of target tissue and distance between microcarrier launch assembly and target tissue (firing distance), for ‘particle gun’ (Bio-Rad) mediated transformation were optimized using embryogenic calli of sugarcane (target tissue), gusA (reporter gene) and hpt (hygromycin resistance gene). Of 12, 18 and 24 µL of DNA (gusA and hpt) suspension used per two bombardments, highest (33.33%) GUS expression was observed with 18 µL of suspension carrying 1.88 µg DNA. Bombardments made into 2-5 mm calli exhibited 55.55% GUS expression, while no expression was obtained in 7.5-12 mm calli after 7 h of their incubation in X-gluc. Out of different firing distances, viz. 2.5, 5.0, 7.5 and 10.0 cm, maximum intensity of GUS expression was recorded when calli were bombarded at a distance of 5.0 cm from the microcarrier launch assembly in the ‘particle gun’.
Page(s): 31-34
ISSN: 0972-5849
Source: IJBT Vol.06(1) [January 2007]

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