Generation of stable recombinant retroviruses containing the β-globin genes linked to complex regulatory elements by using transient transfection

A transient packaging method for a retroviral vector containing β-globin sequences, mediated by Ca₃(PO₄)₂ precipitation, generated stable recombinant β-globin retroviruses with intact viral genomes. We suggest the use of this method as an alternative means to overcome the production of β-globin encoding retroviruses with aberrant viral genomes using the routine packaging procedure.     

In vitro mutagenesis of Xanthomonas campestris α-amylase gene by partially replacing deoxythymidine triphosphate with 5-bromo-2′-deoxyuridine-5′-triphosphate using a PCR technique

Three mutants of the wild type alpha-amylase gene from Xanthomonas campestris pv. campestris 8004 were obtained using a PCR technique in which deoxythymidine triphosphate (dTTP) was partially replaced by 5-bromo-2'-deoxyuridine-5'-triphosphate (BrdUTP), at an optimal dTTP:BrdUTP ratio of 1000:1. Of thre three mutants that were obtained and which were sequenced, one mutant with 40 times higher activity than the wild type alpha-amylase gene product was obtained by using primary PCR products as a template for a second PCR reaction.