Current Issues in Molecular Biology

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In Situ NMR Systems

by Jacqueline V. Shanks

Curr. Issues Mol. Biol. 2001, 3(1), 15-26; https://doi.org/10.21775/cimb.003.015 - 1 Jan 2001

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In situ NMR is becoming an established technology for applications in bioprocessing and metabolic engineering. The in situ NMR biosensor acts as a noninvasive pH, ion, and concentration meter, with ³¹P and ¹³C as the two main isotopes of study. A substantial data base now exists for phosphorus and carbon spectra of bacteria and yeast. In situ NMR can provide many of the state variables needed for modeling glycolytic pathway function. NMR micro-reactor technology has improved significantly in the last decade. Several designs for immobilized cell reactors have been tested, and in particular, considerable gains have been made in the feasibility of studying aerobic, chemostat cultures with in situ NMR. Acquisition of ³¹P spectra from cell suspensions of 3-5% v/v under controlled conditions can be made in 3 - 7 minute time resolution in several systems. Full article

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NMR and Microorganisms

by Jean-Philippe Grivet

Curr. Issues Mol. Biol. 2001, 3(1), 7-14; https://doi.org/10.21775/cimb.003.007 - 1 Jan 2001

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Abstract

This article is an introduction to the use of NMR for the investigation of microbial physiology and metabolism. NMR parameters which determine the sensitivity and resolving power of the method are reviewed. A broad survey of current applications follows. Qualitative uses are described first; they include compound identification and localisation. Quantitative aspects, such as pH, concentration and flux measurements are then examined, as well as the corresponding experimental constraints. The review ends with suggestions of possible future developments in instrument capabilities aimed at improving sensitivity: higher fields, spectroscopic and imaging microprobes. Full article

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Cytoplasmic Expression of Ribozyme in Zebrafish Using a T7 Autogene System

by Kelly Walker, Yuefeng Xie, Yunsheng Li, Qin Zhu, Wenlian Xu, Thomas E. Wagner and Xiaozhuo Chen

Curr. Issues Mol. Biol. 2001, 3(1), 1-6; https://doi.org/10.21775/cimb.003.001 - 1 Jan 2001

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Abstract

A cytoplasmic ribozyme expression system, based on codelivery of a ribozyme vector, a T7 autogene vector, and the T7 RNA polymerase (RNAP), has been developed and used to generate a specific phenotype in zebrafish by targeting a no tail (ntl) mRNA. The expression of the no tail ribozyme sequence is under the control of a tandem of two promoters: The T7 promoter and an adenoviral va 1(pol III) promoter. The coinjection of the ribozyme vector pT7vaRz, the T7 autogene vector pT7T7, and the T7 RNAP resulted in rapid synthesis of the ribozyme against the ntl mRNA in the cytoplasm of the injected zebrafish embryos, generating no tail phenotypes in up to 10 - 20% of the injected embryos. The phenotypic change rates have been found to be related to the concentrations of the plasmid vectors and T7 RNAP injected and to the ratios of the three injected components. This cytoplasmic ribozyme expression system may be useful for efficiently targeting other mRNA and for various biomedical applications. These potential applications may include rapid identification of biological functions of novel genes from zebrafish and humans based on partial gene sequence information and gene therapy of genetic and acquired diseases. Full article

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Curr. Issues Mol. Biol., Volume 3, Issue 1 (January 2001) – 3 articles

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