The Sinorhizobium meliloti ORFeome project cloned 6,314 open reading frames (ORFs) into a modified Gateway entry vector system from which the ORFs could be transferred to destination vectors in vivo via bacterial conjugation. In this work, a reporter gene destination vector, pMK2030, was constructed and used to generate ORF-specific transcriptional fusions to β-glucuronidase (gusA) and green fluorescent protein (gfp) reporter genes. A total of 6,290 ORFs were successfully transferred from the entry vector library into pMK2030. To demonstrate the utility of this system, reporter plasmids corresponding to 30 annotated sugar kinase genes were integrated into the S. meliloti SM1021 and/or SM8530 genome. Expression of these genes was measured using a high-throughput β-glucuronidase assay to track expression on nine different carbon sources. Six ORFs integrated into SM1021 and SM8530 had different basal levels of expression in the two strains. The annotated activities of three other sugar kinases were also confirmed.
- American Society for Microbiology
- Construction and expression of sugar kinase transcriptional gene ...
Construction and expression of sugar kinase transcriptional gene fusions by using the sinorhizobium meliloti orfeome
Courtesy of Courtesy of American Society for Microbiology
A Nitrogen Culinary Experience and How to Remain Safe with Use of Oxygen Monitors
As modernist cuisine has become more popular, restaurant and home chefs alike are turning to liquid nitrogen to create spheres, gels, foams, and even ice cream. While liquid nitrogen can be safely used in a range of culinary applications, there are important safety risks to be aware of when working with this substance. The Hidden Dangers of Liquid Nitrogen in the Kitchen Nitrogen can help chefs freeze alcohol, which doesn`t freeze underfreezer temperatures. Nitrogenalso creates a very rich ice cream, since it...
An Interview with George Hawkins, DC Water
“I have often used the phrase a ‘crisis is a terrible moment to waste.’ Countries and regions that are most impressive on water innovation are usually places that had a crisis of some sort driving the need to innovate.” says George Hawkins, CEO and General Manager of DC Water in an exclusive interview with Rethink Events ahead of next month’s World Water-Tech North America. To view more click here
Resourcing the future 2016, London. Is the Circular Economy strategy on the right track?
The following presentation was given in the form of a speech by Zero Waste Europe Director, Joan Marc Simon at the ‘Resourcing the Future 2016’ conference organised by CIWM in London. Is the Circular Economy strategy on the right track? Yes, but it is still too slow, in need of some fine-tuning and to escape bad habits from the past. The exercise we are undertaking is an ambitious one, close the material loop and turn waste into resources; creating a zero waste society from which the EU’s...
Glyphosate and AMPA in drinking water
Carcinogenic or not? The broad-spectrum herbicide glyphosate is used all over the world in agriculture. Alongside farming, the chemical is also used for weed-killing in domestic gardens and in public and private spaces kept free from «vegetal invasion», such as railway tracks. Glyphosate has been used since the 1970s in pesticides and was hitherto thought to be harmless at typical levels of exposure. However, since the International Agency for Research on Cancer (IARC) – the specialized cancer-re...
Waters and BioanalysisZone are Highlighting our Bioanalytical Chemists
Like the black panther, are analytical scientists going extinct? In a presidential election year, we are reminded of the many tough choices we face—Democrats or Republicans, plastic or paper, ketchup or mustard, soccer or football. And as we watched the Olympic track and field events, for instance, we saw the margin between first and last become more narrow with each subsequent heat. Bioanalysis Zone’s New Investigator’s Award is no different. With a field of 25 outstanding scholars nominated,...