Alpha-Synuclein was expressed in Escherichia coli using plasmid pT7-7 encoding for the protein (courtesy of the Lansbury laboratory, Harvard Medical School, Cambridge, MA). Following transformation, BL21competent cells were grown in LB in the presence of ampicillin (100 mg/ml). Cells were induced with IPTG, cultured at 37 °C for 4 hours and harvested by centrifugation in an Avanti J25 centrifug...

METHODS THAT GENERATE BASE SUBSTITUTIONS ...................................... Fragment Mutagenesis followed by Recombination .................................................. Site Mutagenesis of Intact Genomes ....................................... Incorporation of nucleotide analogues ............................. Bisulfte reaction at single-stranded gaps .....................................

Treatment of human immunodeficiency virus (HIV) type 1 (HIV-1)-infected individuals with antiretroviral (ARV) drugs is highly effective in inhibiting viral replication, increasing both the duration and the quality of life (71). Regimens consisting of combinations of protease (PR), reverse transcriptase (RT), and fusion inhibitors are the current standard of care and typically reduce the circula...

Doramectin is a macrolide antiparasitic that widely used in the treatment of mammalian parasitic diseases. usually produced by Streptomyces avermitilis fermentation using cyclohexanecarboxylic acid (CHC) as precursor; however, growth S. inhibited CHC, resulting low yield doramectin. In this study, high-yielding strain XY-62 was obtained mutant N72 starting strain, then combined with CHC toleran...

In this comparative study, three different mutagenesis kits, namely the MutaGene phagemid in vitro mutagenesis kit (Bio-Rad), the Transformerä Site-Directed mutagenesis kit (Clontech) and the Quik-change site-directed mutagenesis kit (Stratagene) were used for the mutagenesis of IPNS genes. However, a large difference in mutation efficiencies among these kits was encountered. Furthermore, these...

The Rosenberg-Hastings paper (4) defends a model much like that of Foster (1) and argues that the observed mutagenesis contradicts predictions of the amplification model. On the contrary, amplification indirectly causes weak mutagenesis and allows that mutagenesis to have a detectable effect on lac reversion. However, this mutagenesis is a minor side effect that is neither sufficient nor necess...

Insertional mutagenesis screens play an integral part in the annotating of functional data for all sequenced genes in the postgenomic era. Chemical mutagenesis screens are highly efficient but identifying the causative gene can be a laborious task. Other mutagenesis platforms, such as transposable elements, have been successfully applied for insertional mutagenesis screens in both the mouse and...

The T4 mutations px, y and 1206 inactivate an error-prone recombination-like repair system, reducing or abolishing mutagenesis by UV irradiation, MMS, and white light irradiation in the presence of the photosensitizer 8MOP. Both px and y increase some spontaneous mutation rates and slightly enhance proflavin mutagenesis; neither mutation affects thymineless or 2AP mutagenesis appreciably, but b...