Yuri A. Berlin

Incorporation of a pyrene nucleoside analogue into synthetic oligodeoxynucleotides using a nucleoside-like synthon

Abstract A novel phosphoramidite 4 based on the hydroxyprolinol 1 backbone has been synthesized and used to chemically prepare DNA fragments bearing a pyrene-containing nucleoside analogue.

Reagents for Multiple Non-Radioactive Labelling of Oligonucleotides

Abstract Novel phosphoramidite reagents for nonradioactive polylabelling of oligonucleotides have been developed, including two branching reagents, 4 and 8, and a biotin-containing phosphoramidite 19 which can be used to incorporate a biotin moiety into any position of the oligonucleotide.

A Pyrene Seco-Pseudonucleoside in Constructing Interaction-Sensitive Fluorescent DNA Probes

Abstract Based on 4-(1-pyrenyl)-1, 3-butanediol, a pyrene seco-pseudonucleoside, we prepared a nonnucleotide phosphoramidite reagent and a pyrene-modified LCAA-CPG-support and employed them in solid-phase synthesis of pyrene-labeled oligonucleotides with varying numbers and positions of the pseudonucleoside residues. Changes in the fluorescence intensity and the spectral curve shape upon hybridization were observed that depended on the position(s) of the modified unit(s) in the probe.

A high-level prokaryotic expression system: synthesis of human interleukin 1α and its receptor antagonist

Synthetic intronless genes, coding for human interleukin 1 alpha (IL 1 alpha) and interleukin 1 receptor antagonist (IL1ra), have been expressed efficiently in a specially designed prokaryotic vector, pGMCE (a pGEM1 derivative), where the target gene forms the second part of a two-cistron system. The first part of the system is a translation enhancer-containing mini-cistron, whose termination codon overlaps the start codon of the target gene. In the case of the IL1 alpha gene, the high expression level is largely due to the direct efficient translation initiation at the second cistron, whereas with the IL1ra gene in the same system, the proximal translation initiation region (TIR) provides a high level of coupled expression of the target gene. Thus, pGMCE is a potentially versatile vector for direct prokaryotic expression.

SYNTHESIS AND EXPRESSION OF SOME GENES FOR HUMAN CYTOKINES

Abstract Intronless genes for human mature interleukin 1a (IL1a) and its receptor antagonist (IL1ra) have been synthesized and efficiently expressed in a specially devised bacterial plasmid vector as part of a versatile two-cistron prokaryotic expression system.

Construction of a Translation Enhancer-Containing Vector for Gene Expression in a Prokaryotic Two-Cistron System

Abstract A prokaryotic two-cistron system for gene expression, based on the pGEM1 plasmid and containing a translation enhancer in the coding part of the first cistron, has been constructed. The gene to be expressed can be inserted into the vector by means of a PCR-mediated approach using type IIS restriction endonucleases (SDL method). Efficiency of the system is exemplified by the expression of a synthetic gene encoding human interleukin lα.