Factors affecting gene delivery by particle bombardment ofDendrobium orchids

1995 ◽  
Vol 31 (3) ◽  
pp. 131-136 ◽  
Author(s):  
Guo-Ling Nan ◽  
Adelheid R. Kuehnle
1996 ◽  
Vol 96 (4) ◽  
pp. 630-636 ◽  
Author(s):  
Manuel Rey ◽  
Maria Victoria Gonzalez ◽  
Ricardo J. Ordas ◽  
Raffaela Tavazza ◽  
Giorgio Ancora

2012 ◽  
Vol 5 (6) ◽  
pp. 643-654 ◽  
Author(s):  
André Leandro Silva ◽  
Francisco Alexandrino ◽  
Lourena Mafra Verissimo ◽  
Lucymara Fassarella Agnez-Lima ◽  
Lucila Carmem Monte Egito ◽  
...  

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 617a-617
Author(s):  
Hong Y. Yang ◽  
Schuyler S. Korban

Developing an efficient gene transfer system for apple (Malus ×domestica L.) remains a major objective in genetic engineering efforts of this fruit crop. Transient expression of the uidA gene coding for β-glucuronidase (GUS) and driven by the cauliflower mosaic virus 35S promoter (CaMV35S) has been induced in apple cotyledonary explants of mature seeds by tungsten particle bombardment using the Particle Inflow Gun (PIG). Several factors that affect transient expression of the GUS gene in apple cotyledons were investigated. The gene transfer efficiency was monitored by recording the number of blue spots observed on explants two days following bombardment. Precultivation of cotyledons for 18 hours before bombardment significantly increased the number of blue foci. Of the three different precipitation methods tested including water, 25% PEG, and 60% glycerol, the latter was the most effective for coating DNA onto tungsten particles. Washing DNA-coated tungsten particles with 70% ethanol and resuspending in 100% ethanol significantly enhanced gene delivery to cotyledons. The amount of particles used for each bombardment also influenced GUS expression. About 0.5 mg of particles per shot resulted in the highest number of blue foci. Using larger quantity of particles (i.e., 2 mg) drastically decreased GUS expression probably due to the toxicity of tungsten particles.


Gene Therapy ◽  
1999 ◽  
Vol 6 (4) ◽  
pp. 643-650 ◽  
Author(s):  
P R Dash ◽  
M L Read ◽  
L B Barrett ◽  
M A Wolfert ◽  
L W Seymour

1996 ◽  
Vol 96 (4) ◽  
pp. 630-636 ◽  
Author(s):  
Manuel Rey ◽  
Maria Victoria Gonzalez ◽  
Ricardo J. Ordas ◽  
Raffaela Tavazza ◽  
Giorgio Ancora

2014 ◽  
Vol 1060 ◽  
pp. 17-20
Author(s):  
Lalita Leksantikul ◽  
Warayuth Sajomsang ◽  
Theerasak Rojanarata ◽  
Tanasait Ngawhirunpat ◽  
Praneet Opanasopit

The objective of this study was to investigate the transfection efficiency of quaternized chitosan or trimethylated chitosans (TMCs) using the plasmid DNA encoding green fluorescent protein (pEGFP-C2) on human cervical carcinoma cell line (HeLa cells). The factors affecting the transfection efficiency, e.g. degree of quaternization (DQ) and polymer/DNA weight ratio were evaluated. The results revealed that the complexes of TMC30 with DNA had the highest transfection efficiency and safety followed by the complexes of TMC60 and TMC94, respectively. Increasing the DQ of chitosan not only improve the efficiency of gene delivery, but also increase cytotoxicity. In conclusion, TMC30 showed elevated potential as a gene carrier by efficient DNA condensation and mediated the highest level of gene transfection with negligible cytotoxicity in HeLa cells.


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