The Effect Of Sv40 T Antigen On Two Multiple Fission Microalgae Essay

The problem of possible contamination of other long-term cultured tumor cell lines with HeLa cells not only caused an international embarrassment, but also raised the concern of misattributing a specific property so another cell line, for example, a virus or a tumor-specific marker, which actually belongs to HeLa. With the continued and growing use of tissue culture in biochemist research, intra- and interspecific contamination becomes a significant risk. The determination of stable genetic markers on cultured cells is a powerful tool for monitoring such contamination. Recent experiments in which cultured cells and innumerable clones of somatic cell hybrids have been used for genetic analysis have shown that, with the proper use of polymorphic markers to characterize the cells, the possibility of undetected cross contamination of cultures is no longer the problem it once may have

The vital components and techniques of gene cloning are as follows, the DNA sequence that contains the desired gene (EZH2) is amplified by Polymerase chain reaction. PCR was established by Kary Mullis in 1985, popularly known to amplify target sequences of DNA (EZH2) to a billion fold in several hours using thermophilic polymerases (Taq) ,primers and other cofactors (Sambrook and Russell, 2001). Three crucial steps are involved which are Denaturation (at 95°), Annealing of the forward and reverse primers (55-65°) and lastly primer extension (at 72°). After amplification the desired sequence is integrated into the circular vector (pbluescript) forming the recombinant molecule. For the compatibility of the insert and vector, both were digested with (EcoR1) so the same cohesive ends are generated in both, making it easier to ligate. EcoR1 is a restriction enzyme that belongs to the type II endonuclease class which cuts within dsDNA at its recognition site “GAATTC” (Clark 2010; Sambrook and Russell, 2001).

For PEI Transfection, we added 1 ml of Trypsin to a T-25 flask of 293T cells and incubate at 37 0C for 10 minutes. Then, we supplemented 10 ml of DMEM (2.5%), FBS (15), P.S medium to the flack and finally transferred to dish and put back in CO2 incubator. After 5 days, we diluted plasmid DNA (ug) in 250 ul of 150mm NaCl in a sterile tube, and diluted 40 ul of PEI into 250 ul of 150 mm NaCl in second sterile tube. After that, incubated each separate tube for 5 minutes at RT and then mixed the DNA tube with the PEI tube. We put the tube in incubator for 2-5 minutes at RT, supplemented dropwise to cells, and put back into incubator. The important steps that was added helper virus (VSV) to cells and put it in the CO2 incubator; then, collected supernatants that contains our

We tested A type, Alpha type, and mixed type. The key difference between these types are the different genes in Alpha and A type.. These A and alpha type can combine to make a mixed cell via signal transduction pathway. The mixed cells are a combination of A type and Alpha type, these cells can become a haploid, budding haploid, zygote, budding zygote, or a shmoo. A haploid is a single cell. A budding haploid is a single cell with a growth on the side. A zygote is two cells that look like an infinity sign. The budding zygote is 2 cells that are like infinity signs with a growth that looks like a dot. A shmoo looks like a pear, and it the two cells combining together. The mixed type can have shmoos, both haploids, and both zygotes. The A factor and alpha type only have budding haploids and diploids, this is because the A type and Alpha type had nothing to mate with. Single transduction pathways use several steps to produce a cellular response. The yeast cells use G- protein receptors system to mate. G protein receptors are also single transduction pathway. G proteins consist of a signaling molecule, a g protein, G protein coupled receptors and an enzyme. The signaling comes to bind to the G protein on the extracellular side. This causes the G protein coupled receptor to change shape on the cytoplasmic side. When the receptor changes shapes a G protein to bind to it. This activates G

Exploring the Expression of an Auxin Induced Gene pCNT103, a Constitutive Gene GapC and a Cytokinin Induced Cig1 Gene in the DNA and RNA Extracts of Differentiated Shoot, Root, and Callus Tissue of Nicotiana tabacum

Since the HcPro-CA interaction was established using the ZYMV-C. quinoa pathosystem, we first corroborate the possible interaction of HcPro with the A. thaliana homologue of CqCA (AtCA1). Both ZYMV and TuMV HcPro (ZHC and TuHC) were transiently co-expressed with AtCA1 in N. benthamiana. ZYMV and TuMV HC-Pro were fused with YFP (ZHC-YFP and TuHC-YFP respectively) meanwhile AtCA1 was fused with CFP (AtCA1-CFP), proteins localization was monitored by confocal microscopy. As shown in figure XXX AtCA1-CFP co-localized with both ZHC-YFP and TuHC-YFP, displaying the relevance of a relationship between both ZYMV and TuMV HcPro and the A. thaliana homologue of CqCA.

The western blot containing the three different cell lines expressed an equal amount of actin expressed in all three lanes. This served as a control to ensure the plasmids were inserted correctly and there was no interference of any other gene. The lane with cells from mice group 1 showed darker bands on lines corresponding to the proteins Arg as opposed to the lane with mice group 2 cells. The bands corresponding to the protein Abl appeared to be the same on both lanes, demonstrating that Abl was not being accidentally overexpressed as well. These pieces of information elucidated to us that the Arg gene was being successfully overexpressed in line 1. The bands corresponding to Ras-MAPK were darker for mice group 1 than for group 2, demonstrating that this signal transduction pathway for cell proliferation has been downregulated with the overexpression Arg.

One plate was labeled LB/pAMP+kan “+”, one was labeled LB/pAMP+kan “-“, one was labeled LB+, and one was labeled LB-. 100 microliters of the cell suspension from the –pAMP/KAN tube was put on the LB/amp+kan plate using a sterile transfer pipette. Another 100 microliters was added to the LB- plate. The cells were spread evenly over the surface of the plates using a sterile metal spreader. The spreader was sterilized by dipping it in ethanol and flaming it shortly with a Bunsen burner. When the spreader cooled, the cells were evenly distributed on the plate. Using another sterile transfer pipette, 100 microliters of cell suspension from the +pAMP/KAN tube was put on the +LB/amp+kan plate, and another 100 microliters on the LB+ plate. The cells were spread evenly on the plate using the metal spreader previously described. The plates were left to sit for about 10 minutes before being sealed and incubated at 37 degrees Celsius for 24 hours. After 24 hours, the plates were removed from the incubator, and then examined for growth. The number of colonies on each plate were recorded.

15 ml of the chloroplast membrane suspension (CMS) was obtained, and it was put into a clean cold 30 ml beaker. The beaker was kept on an ice. One tube of DCIP and 15 ml of distilled water was obtained. Eight 13 x 100 mm tubes were obtained and each one of them were labeled #1 to #7 and the remaining tube served as a blank. To prepare the blank tube, 0.5 ml of distilled water and 2.5 ml of CMS was added to the tube, covered with parafilm.

Ph.D. Korea Advanced Institute of Science and Technology (KAIST), Biological Science and Engineering (Dewey D. Ryu) (1983)

The plasmid first had to lose S65T-GFP so it could take up yEGFP. This was done by adding Pac1 and Asc1 to the plasmid to cut out the gene and leave the proper sticky ends for yEGFP to ligate to. A gel electrophoresis was then used to separate pWDH444 and S65T-GFP. The pWDH444 was cut from the gel electrophoresis and removed from the gel. After successful removal of the plasmid from the gel pWDH444 and yEGFP were ligated to create a new plasmid with the ideal GFP. The new plasmid, pWDH444-yEGFP was the ideal plasmid for yeast due to the enhanced GFP with a codon bias for yeast and it had double the light output of S65T-GFP. The difference in fluorescence was detected using a extra sensitive fluorimeter than normal and 495-nm glass, sharp cut-off filter. The filter reduced the amount of light shattering leading to an improved sensitive reading from the

Dr. Alwine is a Professor of Cancer Biology and an investigator in the Abramson Family Cancer Research Institute in University of Pennsylvania. He studies DNA viruses and how they react with the consequences of inducing cellular stress responses. Dr. Alwine received his Bachelors of Science in Chemistry from Elizabethtown College in 1969 and his Ph. D in Biological Chemistry from Pennsylvania State University in 1974. He began his interest in research with Simian virus 40 where he developed the Northern Blot technique, also known as the RNA blotting technique, at a laboratory in George Stark in the Biochemistry Department of Stanford University. It was named after the Southern Blot technique, which blots for DNA, and the Western Blot technique, which blots for proteins. It is not commonly used for clinical or diagnostic purpose but it is mainly used in research.

Chlamydomonas is a biflagellated green algae consisting of one circular cell only. Gametes in theirs structure are identical in chlamydomonas. The distinctive feature of Chlamydomonas is an extensive chloroplast, which has the pyrenoid, aiding in starching the formation. Not only that, but also it posses the capability of asexual and sexual reproduction. Single-celled algae such as Chlamydomonas produce daughter protoplasts during the mitosis stage. Each new cell has a cell wall, which forms around it, so the temporary colonies emerge. These colonies release zoospores, so-called daughter cells. During the sexual reproduction process, two vegetative cells play the roles of male and female gametes. In this case, a diploid zygote forms.

In order to test FDH protein (O59078) expression, the recombinant vector was transformed into the BL-21-(DE3)-RIL strain. Ampicillin was used as a selection marker. Transformed single colony was inoculated in Luria-Bertani (LB) medium with Ampicillin and grown overnight at 37C. 1% of the

WP1. Starting point of the project is the molecular analysis of Arabidopsis candidate DNA insertion lines (obtained from the NASC stock center) harbouring T-DNA insertions in the gene encoding HMGA. Typically seeds of a segregating population are