Selection Marker positive

Clostridium spp. used for exemplification Wild type or mutant Negative selection marker Positive selection marker Selection criteria Plasmid example Principle genes inactivated/added References... 

Fig. 3 Measurement of signal stored in the memory buffer. Two markers can be set to arbitrary selected delay positions along the buflfer length and signal values can be read and compared.

Figure 19 Sample gel of the results of a PCR. Lane 1 is a 100-bp molecular marker lanes 2-6 are samples. The presence of the top bands (the species-specific endogenous gene) demonstrates that the PCR amplification was successfiil. Lack of the middle band (the introduced effect gene) and the bottom band (the selectable marker gene) in lane 3 indicates that sample is negative for the effect gene. Presence of all three bands in the remaining lanes indicates the samples are positive for the effect gene...

The plant sample in lane 6 is also positive for the transgene of interest. Because the band for the effect gene (middle band) is typically fainter than the band for the selectable marker gene (bottom band), it appears that for lane 6, the PCR product amplitication for the effect gene is below the assay detection threshold. Because the selectable marker is clearly present and the PCR amplitication worked, lane 6 can be interpreted as a positive result for the transgene of interest. 

Fig. 3. Representation of (A) the two basic types of targeting vectors, (B) positive and negative selection markers, (C) the hit-and-run approach to introduce subtle mutations, and (D) the CRE/loxP recombinase system.

Homologous recombination occurs approximately 1000-fold less frequently than non-homologous recombination, therefore methods have been developed to enrich for homologous recombination events. A widely applied method includes the use of a positive and a negative selection marker and does not require the expression of the target gene in ES cells (Mansour et al., 1988). The targeting construct is based on a replacement-type vector... 

Targeting vector A vector containing a positive selection marker (e.g., neomycin-resistance gene), negative selection marker (thymidine kinase gene) and a multiple cloning site. 

Fig. 2. Mutagenesis strategy. A neo gene replaced one-third of the ORF and was used as a positive selectable marker. The HSV thymidine gene (HSV-ffc) was used for negative selection. The vector was linearized with Xbal digestion. The targeted events were screened by Southern blot. Two Xbal sites are in the outside of the targeting vector (the 3 -end one is just at the end of the vector, which cannot be digested by Xbal if the vector is randomly inserted). An outside probe is located at the 5 -end. The genotype of the mice can be easily identified based on the size of the hybridized bands, 6.7 kb (mutant) vs 6 kb (wild-type).

Mapview has many useful options, which are well described in the oifline help. Some maps have more than one tier, each displaying different types of markers, such as markers positioned with varying confidence thresholds on a linkage or radiation hybrid map. It is possible to zoom in and out, highlight markers across maps, color code different tiers, display markers using different aliases, change the relative position of the displayed maps, and search for specific markers. To retrieve additional information on a marker from any of the maps, double-click on its name to perform a Simple Search (as described above). A separate browser window will then display the GDB entry for the selected marker. 

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