A simple and rapid method for generating a deletion by PCR

Polypeptide binding specificities of Saccharomyces cerevisiae oligosaccharyltransferase accessory proteins Ost3p and Ost6p

“…Variants with amino acid replacements (pMBP-OST6L K96Q,K99Q and pMBP-OST3L Q103K,Q106K ) were constructed as described by Imai et al 35 For Gas1p expression in E. coli, the DNA sequence corresponding to mature Gas1p (from the signal peptidase cleavage site to the GPI-anchor site) as predicted in the UniProt Knowledgebase: residues 23-528 of Gas1p (P22146, YMR307W), was cloned into a derivative of the pMAL-p2x vector, using PCR primers incorporating BamHI and HindIII restriction sites, giving pMBP-GAS1.…”

Calmodulin-independent, agonistic properties of a peptide containing the calmodulin binding site of estrogen receptor α

“…PCR, used for that purpose (Imai et al, 1991), was performed using Expand Long Template PCR A c c e p t e d M a n u s c r i p t 8 System (Roche Applied Science) with C-terminal 5' -gcttggccaaaggttggcagc -3' and N-termina l 5' -gccgaccagatggtcagtgcc -3' primers. PCR product was purified using…”

“…Wizard SV Gel and PCR Clean-Up System (Promega) and self-ligated (Imai et al, 1991) after blunt-ending with Klenow fragment.…”

Identification of a p130 domain mediating interactions with cyclin A/cdk 2 and cyclin E/cdk 2 complexes

“…These three proteins, pRB, p107 and p130, were ®rst described as targets of the adenovirus E1A oncoprotein which forms a complex with each of the RB-related proteins resulting in disruption of their normal function and deregulation of cellular growth control (reviewed in Whyte, 1995). Oncoproteins from other DNA tumor viruses including the SV40 large T antigen and HPV E7 proteins have been shown to target the RB-related proteins in a manner analogous to E1A (DeCaprio et al, 1988;Whyte et al, 1988;Dyson et al, 1989aDyson et al, , 1990Ludlow et al, 1990;Imai et al, 1991;Davies et al, 1993;Wolf et al, 1995). These observations ®rst suggested that the RB-related proteins (pRB, p107 and p130) control key checkpoints in cellular growth control.…”

“…In almost all cases involving interactions with pRB, phosphorylation of pRB abrogates the interaction. For example, phosphorylation of pRB results in its dissociation from E2F transcription factors (Chellappan et al, 1991;Helin et al, 1992;Shan et al, 1992;Ewen et al, 1993;Dynlacht et al, 1994;Bremner et al, 1995;Suzuki-Takahashi et al, 1995) and from E1A, T antigen and E7 (Ludlow et al, 1990;Imai et al, 1991;Templeton and Weinberg, 1991). Phosphorylation of p107 appears to regulate its association with E2F proteins in a manner analogous to pRB (Bejersbergen et al, 1995;Xiao et al, 1996).…”