Description:
Information about the Arabidopsis project (Moderated)
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At gene expression in E.coli
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Hello, I ever tried to express a few At transcription factor (TF) genes in E.coli using the Novagen pET32a(+) vector and BL21(DE3) as well as Rosetta-gami strain. Only one of them (aa shorter than the others) was successfully expressed in the Rosetta-gami strain, suggesting that codon bias is possibly a problem for plant TFs exp in E.coli. However, the sole expressed TF was not in the soluble fraction; it is in the form of inclusion body, although I induced it at 30C for only 3 h. It takes a lot effort to get it out from the inclusion body and then refold it. So far, I did not see much use of using E.coli to express a TF protein. Another lab mate also got an unknown protein gene expressed in the inclusion body. A friend saw a fungal gene expressed in the inclusion body too. I ever read a paper describing a strategy in E.coli(not too sure) that adds a tag to target protein and secrete the protein into the medium, is any company selling this type of vector? What made I very doubtful is I saw a published paper expressing one of the TF genes I failed to get exp in E.coli and they succeeded with BL21(DE3) strain. I checked their "Materials and methods" section and found that the IPTG concentration they used was also 1 mM, the same as I ever used. Other than this, no more detail was included in their paper; they just saw "according to the manufacturer's manual". No idea whether that TF was expressed in soluble form or in inclusion body. No info was included in the supplemental data. Were they cheating me?... more »
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software for scanning protein localization.
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Hello all Does someone know some software which could scan the exact localization of expression protein in protoplast of Arabidopsis? I have taken some pictures when I saw the expression within the protoplast, and the protein was expressing near the membrane of the cell. I heard that there is a soft ware which can scan the exact position of the expression,... more »
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Cvi Cosmid library
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Hello I am looking for a genomic library for Arabidopsis ecotype Cvi in a binary Cosmid vector. If someone somewhere has made it and can share it for research purpose it would be great. Otherwise any suggestion on which Binary Cosmid vector should I use? looking forward for responses thanks Jagreet ... more »
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pseudogene or not
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Hi collegues, I recently cloned some genes of Brassica napus by RT-PCR/RACE through mining ESTs in GenBank. The primers were designed based on ESTs. After sequencing more than 2 clones for each gene, I found, for quite a few genes, obviously, alleles exist. The most common is nucleotide substitution, next indel(insertion/deletion). An interesting phenomenon I found is for those alleles with a a long insertion compared to the other allele, the insertion is not 3-folds, i.e. the insertion leads to premature stop codon in the insertion or immedialtely after the insertion. The means that the translated amino acid sequence of one allele is normal (compared to At homolog), while the other is much shorter (premature stop codon).... more »
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help
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Dear all Now I come across with a problem about my study. When I overexpress the gene which i am interested in and get the transgenic plant.Then I find that some lines have the phenotype which is sterile. However I do not know why this overexpression can cause sterile. Then I analyze the expression pattern of the promoter. But it does not express neither in pollen nor pollen tube. It expresses in guard cells of stoma and style.... more »
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cis element analysis - using co-expressed genes.
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Dear Jianling, If you're interested in promoters of Arabidopsis genes... we have developed a site calculating the correlation of expression patterns for Arabidopsis genes from a database of Affymetrix microarray experiments. Co-expressed genes are often co-regulated and therefore analysis of a set of promoters for strongly co-expressed genes (including the promoter... more »
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UNC Assistant Professor Position
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ASSISTANT PROFESSOR QUANTITATIVE BIOLOGY OR BIOINFORMATICS DEPARTMENT OF BIOLOGY UNIVERSITY OF NORTH CAROLINA AT CHAPEL HILL The Department of Biology at the University of North Carolina at Chapel Hill invites applications for a tenure-track Assistant Professor in Quantitative Biology or Bioinformatics. We seek a researcher who is applying quantitative and/or... more »
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Postdoc and PhD student positions available
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Postdoc and PhD student positions are available at the Institute of Botany, University of Basel, Switzerland, to study the relationship between viruses and the plant RNA silencing machinery. The project is funded by Swiss National Science Foundation. We are looking for a motivated and hard working young scientist with a solid background in molecular plant virology, basic skills in biochemistry, experience with RNA work including small RNA blot hybridization, and a deep interest in RNA silencing and viruses.... more »
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Join the Fastest Growing Bio Community
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Join the Fastest Growing Bio Community - you know Facebook, Xing or MySpace, LinkedIn. In biology it exists as well: Visit academic portal [link] register, confirm registration in email - join group, institute, create own group of interesst and share data, contacts, micorscope images, build network.... more »
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