Systems for Plant Protoplast Transformation

1983 ◽  
pp. 57-60 ◽  
Author(s):  
T. Manzara ◽  
P. F. Lurquin
Keyword(s):  
Protoplast Transformation ◽  
Plant Protoplast

Approaches to Plant Protoplast Transformation Using Drug Resistance and Auxotroph Complementation as Selective Markers

1983 ◽  
pp. 265-276 ◽  
Author(s):  
R. D. Shillito ◽  
G. Lazar ◽  
J. Paszkowski ◽  
K. Shimamoto ◽  
Z. Nicola-Koukolikova ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Protoplast Transformation ◽  
Plant Protoplast

Enhanced polyethylene glycol (PEG)–mediated protoplast transformation system for the phytopathogenic fungus, Ganoderma boninense

2021 ◽  
Author(s):  
Fook-Hwa Lim ◽  
Omar Abd Rasid ◽  
Abu Seman Idris ◽  
Abdul Wahab Mohd As’wad ◽  
Ganesan Vadamalai ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Phytopathogenic Fungus ◽  
Ganoderma Boninense ◽  
Protoplast Transformation ◽  
Transformation System

scholarly journals EXPRESSION OF b-XYLOSIDASE ENCODING GENE IN PHIS/ Bacillus megaterium MS SYSTEM

2015 ◽  
Vol 2 (1) ◽  
pp. 25 ◽  
Author(s):  
Sri Sumarsih
Keyword(s):  
Culture Medium ◽  
Culture Supernatant ◽  
Specific Activity ◽  
Agar Medium ◽  
Nitrilotriacetic Acid ◽  
Protoplast Transformation ◽  
E Coli ◽  
Recombinant Cells ◽  
Relative Molecular Weight ◽  
Encoding Gene

b-Xylosidase encoding gene from G. thermoleovorans IT-08 had been expressed in the pHIS1525/ B. megaterium MS941 system. The b-xylosidase gene (xyl) was inserted into plasmid pHIS1525 and propagated in E. coli DH10b. The recombinant plasmid was transformed into B. megaterium MS941 by protoplast transformation. Transformants were selected by growing the recombinant cells on solid LB medium containing tetracycline (10 µg/ ml). The expression of the b-xylosidase gene was assayed by overlaid the recombinant B. megaterium MS941 cell with agar medium containing 0.2% ethylumbelliferyl-b-D-xyloside (MUX). This research showed that the b-xylosidase gene was succesfully sub-cloned in pHIS1525 system and expressed by the recombinant B. megaterium MS941. Theaddition of 0.5% xylose into the culture medium could increase the activity of recombinantactivity of recombinant of recombinantb-xylosidase by 2.74 fold. The recombinant B. megaterium MS941 secreted 75.56% of the expressed b-xylosidase into culture medium. The crude extract b-xylosidase showed the optimum activity at 50° C and pH 6. The recombinant b-xylosidase was purified from culture supernatant by affinity chromatographic method using agarose containing Ni-NTA (Nickel-Nitrilotriacetic acid). The pure b-xylosidase showed a specific activity of 10.06 Unit/mg protein and relative molecular weight ± 58 kDa.

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