Vectors expression test

Whether rAAV distributes to sperm cells has been of significant concern because of the possibility of introducing mutations in sperm. While AAV DNA can be found in the testes of rodents, rabbits, and dogs following either hepatic arterial or IM injections, these signals appear to be localized to the testis basement membrane and the interstitial space, with no intracellular signal observed. In mice and rats, there was a dose-dependent increase of vector expression in gona-... 

To determine whether other glycoproteins in the US2-US11 region affected the class II pathway. His 16 cells were infected with Ad vectors expressing US2, US3, US7, US8, US9, US 10, or USl 1 and tested in CD4 T cell assays. The CD4 T cell clones were specific for a tuberculosis antigen, Mtb39, and this protein was added... 

Kim TH, Kim HJ, Park JS, Kim Y, Kim P, Lee HS (2005) Functional analysis of sigH expression in Corynebacterium glutamicum. Biochem Biophys Res Commun 331 1542-1547 Knoppova M, Phensaijai M, Veseiy M, Zemanovd M, Nesvera J, Patek M (2007) Plasmid vectors for testing in vivo promoter activities in Corynebacterium glutamicum and Rhodococcus erythropolis. Curr Microbiol 55 234-239... 

The dot blot will provide a qualitative estimate of antibody titer (Protocol 21.3). The same test can be done quantitatively by ELISA (enzyme-linked immunosorbent assay), but the technically simpler dot blot gives adequate information, uses less protein, and does not require an ELISA plate reader. If the same protein has been expressed in two different fusion-protein vectors, the test should be performed with the protein not used for immunization. If there is only one fusion protein, e.g., a GST-fusion, then a second dot blot can be done in parallel with just GST alone to compare the titers. If the antiserum recognizes the protein, there should be a lower titer of antibodies against GST alone as compared to against the entire fusion protein. A preimmune control is always advisable. Figure 21.1 shows a sample dot blot from a monoclonal antibody screen. 

Some Basics. The field theory of electrostatics expresses experimentally observable action-at-a-distance phenomena between electrical charges in terms of the vector electric field E (r, t), which is a function of position r and time t. Accordingly, the electric field is often interpreted as force per unit charge. Thus, the force exerted on a test charge q, by this electric field is qtE. The electric field due to a point charge q in a dielectric medium placed at the origin r = 0 of a spherical coordinate system is... 

Compared with whole plants, there has been limited development of foreign protein expression systems specifically for use in tissue culture. Some modifications of expression constructs have resulted in improved protein accumulation or have allowed simplified protein recovery. However, in general, modified expression systems have been tested only in a restricted number of cases and have not resulted in the large increases in product yield required for plant cultures to compete with other foreign protein production vehicles. Transient expression techniques, for example using viral vectors, that have been developed for use in whole plants have not yet been applied in plant tissue culture. 

Wigdorovitz et al. [39] used a TMV expression vector to produce VP1, the 26-kDa structural protein from FMDV, and tested it in mice. Mice injected intraperitoneally with leaf extracts prepared from infected plants mounted an antibody response against the plant-derived protein. All immunized mice were protected when challenged with virulent FMDV. One-year old calves immunized with plant extracts containing VP1 also developed FMDV-specific antibody responses. 

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