1887

Abstract

Transcripts of the gas vesicle genes and were detected in single filaments of the cyanobacterium using reverse transcription and quantitative real-time PCR. Primers were designed to amplify short sequences within and three length variants of . With genomic template DNA, and using Sybr Green to monitor product accumulation, similar amplification efficiencies were observed for each of these genes. The relative copy numbers of length variants in genomic DNA from five gas vesicle genotypes determined by real-time PCR were similar to those indicated by sequencing the gas vesicle gene clusters. The precipitation of cDNA reverse-transcribed from cellular RNA from single filaments was required before amplification of the gene fragments; without this step it was not possible to detect the accumulation of the expected amplicons by dissociation analysis. Precipitation was also necessary to ensure the generation of product curves that allowed linear regression in an early stage of PCR, a prerequisite for the quantification of low-input cDNA amounts without the need for standard curves. This report shows that different length variants are transcribed within single filaments, both from laboratory cultures and from natural samples taken from Lake Zürich. This has implications for the efficiency of buoyancy provision by the possible production of gas vesicles of different strengths within individual cyanobacterial filaments. The hypothesis that post-transcriptional regulation may influence the type of protein (GvpC) present in gas vesicles is presented.

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2005-01-01
2019-10-17
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