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91.
In this work it is proposed that in evolution amino acid substitutions implying strong physico chemical and structural differences are more relevant and more frequent than substitutions between similar amino acids. This analysis is made over a group of protein families representing about 10 000 substitutions and as examples the evolutionary trees of fibrinopeptides A and calcitonins were constructed and compared. 相似文献
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The effect of incubation temperature on the background staining of Western blots with monoclonal antibodies to a human milk protein, alpha-lactalbumin (Mr 14,500), is presented. Human milk proteins were electrophoretically separated and transferred to nitrocellulose membranes which were then blocked with bovine serum albumin, "BLOTTO", casein, or Tween 20. They were subsequently incubated with mouse monoclonal antibody to human alpha-lactalbumin, biotinylated anti-mouse antibody, strepavidin-biotinylated horseradish peroxidase complexes and a substrate containing diaminobenzidine and nickel chloride. Reduction of incubation temperature from 37 degrees C to 22 degrees C and 4 degrees C was found to decrease the extent of non-specific background staining independent of the type of blocking reagent used. Good specific staining with minimal background was found using 0.1% Tween 20 in phosphate-buffered saline, pH 7.2, as blocking agent and incubation temperatures of 4 degrees C. 相似文献
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Pearlyn J Y Toh Jason K H Lai Anke Hermann Olivier Destaing Michael P Sheetz Marius Sudol Timothy E Saunders 《EMBO reports》2022,23(9)
YAP, an effector of the Hippo signalling pathway, promotes organ growth and regeneration. Prolonged YAP activation results in uncontrolled proliferation and cancer. Therefore, exogenous regulation of YAP activity has potential translational applications. We present a versatile optogenetic construct (optoYAP) for manipulating YAP localisation, and consequently its activity and function. We attach a LOV2 domain that photocages a nuclear localisation signal (NLS) to the N‐terminus of YAP. In 488 nm light, the LOV2 domain unfolds, exposing the NLS, which shuttles optoYAP into the nucleus. Nuclear import of optoYAP is reversible and tuneable by light intensity. In cell culture, activated optoYAP promotes YAP target gene expression and cell proliferation. Similarly, optofYap can be used in zebrafish embryos to modulate target genes. We demonstrate that optoYAP can override a cell''s response to substrate stiffness to generate anchorage‐independent growth. OptoYAP is functional in both cell culture and in vivo, providing a powerful tool to address basic research questions and therapeutic applications in regeneration and disease. 相似文献
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Cheng Hong Toh Kuo-Chen Wei Chen-Nen Chang Shu-Hang Ng Ho-Fai Wong Ching-Po Lin 《PloS one》2014,9(10)