What is GV3101?
The GV3101 Ti plasmid has the T-DNA region sequences deleted and transformation with a binary vector containing the missing T-region results in a functional T-DNA binary system that allows for transfer of genetic material into a host plant’s genome.
What is EHA105?
The EHA105 strain is useful for transgenic operations of rice, tobacco and other plants. EHA105 contains a rifampicin resistance gene (rif). EHA105 strain also contains an amber basic Ti plasmid pEHA105 (pTiBo542DT-DNA) without self-transport function, which contains the vir gene.
What is Agrobacterium strain?
An Agrobacterium strain is determined by the chromosomal background, known chromosomal mutations herein, and the type of helper plasmid(s) present. In many cases, strain names refer to the combination of a particular chromosomal background with a certain helper plasmid.
How do you make Agrobacterium competent cells?
Competent cells are made by streaking the master stock onto a rifampicin plate, growing a single colony in a large culture, and then washing the pellet of cells several times with 10% glycerol and flash freezing in liquid nitrogen.
How do you make Acetosyringone stock solution?
Acetosyringone (AS) – 100mM Stock (mw. 196.20 g/mol): • Dissolve in 95% ethanol and adjust volume with water. Dissolve 0.3924 g in 12 ml 95% ethanol, then add 8 ml of sterile milli-Q water to equal 20 ml. Filter Sterilize and store at -20°C.
What is disarmed Ti plasmid?
Abstract. Agrobacterium-mediated plant transformation has been used widely, but there are plants that are recalcitrant to this type of transformation. This transformation method uses bacterial strains harboring a modified tumor-inducing (Ti) plasmid that lacks the transfer DNA (T-DNA) region (disarmed Ti plasmid).
Which bacteria is responsible for crown gall?
Crown gall is caused by the bacterial plant pathogen, Agrobacterium tumefaciens. Crown gall bacteria enter plant roots through wounds.
What is electroporation method?
Electroporation is a physical transfection method that uses an electrical pulse to create temporary pores in cell membranes through which substances like nucleic acids can pass into cells.
How long does Agrobacterium take to grow?
A. tumefaciens grows optimally at 28°C. The doubling time can range from 2.5–4h depending on the media, culture format and level of aeration….Induction Broth (IB medium) (Winans et al., 1988)
| 1X IB Medium | volume/L | Final conc (1x) |
|---|---|---|
| 50% glucose | 10 ml | 0.5% (28 mM) |
| 0.1 M phosphate buffer (pH 7.0) | 500 μl | 50 μM |
Can Acetosyringone be autoclaved?
Acetosyringone is a naturally occurring compound secreted from wounded dicot plant tissues and is a chemotractant for Agrobacterium sp. It is generally used at 5 uM to 200 uM to enhance transformation in monocots. Acetosyringone should NOT be autoclaved. This is a sterile solution of Acetosyringone in DMSO.
How do you store Acetosyringone?
Storage in DMSO for up to 12 months at room temp (I have found other sources that say to keep stored at 4C). If you have prepared more dilute stocks, those should be stored at -20C for no more than a month.
What is the function of Ti plasmid?
A key feature of Ti plasmids is their ability to drive the production of opines, which are derivatives of various amino acids or sugar phosphates, in host plant cells. These opines can then be used as a nutrient for the infecting bacteria, which catabolizes the respective opines using genes encoded in the Ti plasmid.
What is the chromosomal background of the gv3101 strain?
The GV3101 strain has a C58 chromosomal background with rifampicin resistance and the Ti plasmid pmp90 (pTiC58DT-DNA) with gentamicin resistance.
What is type gv3101 Agrobacterium tumefaciens?
Type Agrobacterium tumefaciens. GV3101 Agrobacterium Strain Description. GV3101 strain is C58 background, containing screening label, rifampicin resistant gene RIF in nuclear genes, in order to facilitate the transformation of the operation, the strains carrying nopaline type Ti plasmid pMP90 without its transport function (pTiC58DT-DNA),
What is the genotype of gv3101 chemically competent cell?
The genotype of GV3101 Chemically Competent Cell is C58 (rif R) Ti pMP90 (pTiC58DT-DNA) (gent R) Nopaline. Background of GV3101 strain is C58. It contains rifampicin resistant gene (rif) in nuclear genes as screening label. GV3101 carries nopaline type Ti plasmid pMP90 (pTiC58DT-DNA) to facilitate transformation.
What is the gv3101 Ti plasmid?
The GV3101 Ti plasmid has the T-DNA region sequences deleted and transformation with a binary vector containing the missing T-region results in a functional T-DNA binary system that allows for transfer of genetic material into a host plant’s genome.