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GV3101 (pSoup-p19) Competent Cell


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GV3101 (pSoup-p19) Competent Cell

Cat#
ACC-102
Product Name
GV3101 (pSoup-p19) Competent Cell
Product Overview
The genotype of GV3101 (pSoup-p19) Competent Cell is C58 (rifR) Ti pMP90 (pTiC58DT-DNA) (gentR) Nopaline(pSoup-p19-tetR). The p19 protein is derived from tomato bush dwarf virus, which can inhibit the RNA silencing effect of foreign genes, improve the stability of heterologous gene transcripts, and promote the expression of heterologous proteins. Therefore, it is widely used in transient expression system in transgenic plants and tobacco leaves, Arabidopsis leaves, tomato leaves or protoplasts. The background of the GV3101 (pSoup) strain is C58 type, and the nuclear gene of GV3101 contains a screening tag - rifampicin resistance gene (rif). In order to facilitate the transformation, the GV3101 strain carries a nopaline-based Ti plasmid pMP90 (pTiC58DT-DNA) without self-transport function, which contains the vir gene. The vir gene is an essential component for the insertion of T-DNA into the plant genome, and the T-DNA transfer function of the pMP90 (pTiC58DT-DNA) plasmid itself is disrupted, but the foreign binary vector T-DNA can be transferred smoothly. Since the pMP90 (pTiC58DT-DNA) plasmid contains the screening tag: gent, the GV3101 strain has gentamicin resistance. Transferring the pSoup-p19 plasmid into the GV3101 strain is the GV3101 (pSoup-p19) strain, which helps the pGreen, 62SK, pGs2 series plasmids to replicate in Agrobacterium and helps the GV3101 (pSoup-p19) strain to have tetracycline (tet) resistance.
Applications
GV3101 (pSoup-p19) Competent Cell is suitable for transgenic operations of Arabidopsis, tobacco, corn, potatoes and other plants.
Notes
1. The volume of the added plasmid should not be greater than 1/10 of the competent volume; When the plasmid is impure or contaminated with organic substances such as ethanol, the conversion efficiency drops sharply. 2. Gently handle when mixing in the plasmid. If a high concentration of plasmid is converted, the amount of bacteria ultimately used for plating can be reduced accordingly. GV3101 (pSoup-p19) chemically transformed competent cells have tetracycline resistance, but when screening positive clones after transfer into the target plasmid, it is necessary to add antibiotics resistant to the target plasmid without tetracycline. 3. When the density of positive clones on the plate is too large, the positive clones grow slower and the colonies become smaller due to insufficient nutrition. In order to obtain large colonies, the amount of plasmid should be reduced. 4. The concentration of rifampicin should not be higher than 25 μg/ml. The excessive concentration of rifampicin is not conducive to the growth of Agrobacterium and will reduce its growth rate and transformation efficiency. 5. The purpose of adding rifampicin to the culture medium is to prevent the growth of bacteria and to screen Agrobacterium. According to the resistance of the strain used, the addition of streptomycin or gentamicin can prevent the loss of Ti plasmid, but these antibiotics is not conducive to the transgenic operation of Agrobacterium. Therefore, in general, Agrobacterium is not added with streptomycin or gentamicin, and the probability of loss of Ti plasmid is extremely low (can be ignored).
Storage
Store at - 80 centigrade for 12 months.
Kit Components
GV3101(pSoup-p19): 100μl/tube * 10 tube/50 tube/100 tube; pGs2(control vector, 10ng/μl): 10μl.
Sensitivity
The conversion efficiency of pGs2 (kanamycin resistance) plasmid in GV3101 (pSoup-p19) Competent Cell is >10^3 cfu/μg DNA.
Sample Type
Arabidopsis, tobacco, corn, potatoes and other plants
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