[ccp4bb] Fwd: [ccpem] Glutamate receptor cryo-EM at the MRC LMB

2021-06-16 Thread Krieger, James M
Hi everyone, I’m forwarding this position in structural biology and drug development at the LMB in Cambridge with my former PhD supervisor, Ingo Greger, in collaboration w AstraZeneca in case anyone is interested who is not on CCPEM. It’s a very dynamic, interdisciplinary group that I highly re

[ccp4bb] Bioinformatician position at PDBe

2021-06-16 Thread John Berrisford
Dear Colleagues, We have a bioinformatician position available in the PDBe team at the European Bioinformatics Institute (EBI) on the Wellcome Genome Campus near Cambridge. We are looking for a scientific programmer/bioinformatician with a strong structural biology background and software e

[ccp4bb] aimless scale without merging data running Fail

2021-06-16 Thread Jiang Xu
Hello guys, I am having some problems running Aimless from the CCP4i packages. I want to scale without merging the data. The input mtz file for aimless was either the mtz file directly from Imosflm integration, or from Pointless from a previous run on Imosflm. I ran both pointless and aimless su

Re: [ccp4bb] aimless scale without merging data running Fail

2021-06-16 Thread Phil Evans
I think it’s trying to write a file TILEIMAGE.img to a directory where you aren’t allowed to write. I can’t remember what is done in ccp4i - I believe it should work ok in ccp4i2, which produces better reports, and is generally recommended as a replacement for ccp4i As a workaround, you might b

Re: [ccp4bb] aimless scale without merging data running Fail

2021-06-16 Thread Jiang Xu
Hello Phil, Thank you for the comments. I noticed that the ccp4 package I used was 7.0. After upgrading to the latest version, the problem was solved. I also have another question about Aimless. I want to use aimless to scale without merging the data for generating the "table1" of my data. I

[ccp4bb] Looking for proteins for undergraduate biochemistry lab

2021-06-16 Thread P. H
Hello All, We are looking for some candidate proteins for an undergraduate level advanced biochemistry lab. They should be expressed in bacteria, simple enough to purify and it will be nice to perform some simple characterization experiments(binding assays, enzymatic assays). Any suggestions? Tha

Re: [ccp4bb] aimless scale without merging data running Fail

2021-06-16 Thread Jiang Xu
Hello Phil, I just read my email and found I may not ask the question clearly. So my question is, should I use the integrated data from Imosflm, or should I use the output from Pointless as the input of Aimless to do scale without merging the data? Thank you, Best, Jiang Lin Chen Research Group

Re: [ccp4bb] Looking for proteins for undergraduate biochemistry lab

2021-06-16 Thread Chun Luo
Many phosphatases, such as lambda phosphatase, have good soluble expression in E. coli. Their activity can be shown by simply colorimetric assay. From: CCP4 bulletin board On Behalf Of P. H Sent: Wednesday, June 16, 2021 3:19 PM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Looking for protein

Re: [ccp4bb] Looking for proteins for undergraduate biochemistry lab

2021-06-16 Thread Roger Rowlett
Human carbonic anhdydrase II is very expressible in *E. coli,* and purifiable in one step via affinity chromatography with para-aminobenzenesulfonamide affinity resin (which is relatively easy to make, and reusable for many years.) It can be assayed by stopped-flow spectrophotometry for CO2 hydrati

Re: [ccp4bb] Looking for proteins for undergraduate biochemistry lab

2021-06-16 Thread Peat, Tom (Manufacturing, Parkville)
I agree with Roger that human CAII is easy to express in E. coli at high level and does not need a tag for purification- and that it is a good lesson for students to purify proteins without tags (they sometimes have better activity and crystallise better without having the tag). It is also easy

Re: [ccp4bb] Looking for proteins for undergraduate biochemistry lab

2021-06-16 Thread Brian Shoichet
FWIW, many of the beta-lactamases fall into the same category: single step purification on an affinity resin, a few litres can give you 100 mg quantities, great spectrophotometric assay, several good substrates and many inhibitors, both covalent (teaching kinetics) and non-covalent. brian On Wed,

Re: [ccp4bb] aimless scale without merging data running Fail

2021-06-16 Thread Phil Evans
Dear Jiang If you gave Mosflm the “correct” space group, then you can skip the Pointless step, but I don’t think the pipelines in ccp4i and certainly ccp4i2 let you do that. However, it shouldn’t hurt. I don’t think you can skip the merging step in Aimless, and anyway it is that step which ge