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10.1007/s11033-020-05717-8 http://scihub22266oqcxt.onion/10.1007/s11033-020-05717-8 32772343!7415195!32772343     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 243.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Mol+Biol+Rep 2020 ; 47 (9): 7283-7289 Nephropedia Template TP {{tp|p=32772343|t=2020. Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1) |pdf=|usr=}}{{32772343}} gab.com Text Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1) JO: Mol Biol Rep http://www.kidney.de/pget.php?&tw=1&pmid=32772343 #FOAvip It is well established that Escherichia coli represents a powerful tool for the over-expression of human proteins for structure/function studies. In many cases, such as for membrane transporters, the bacterial toxicity or the aggregation of the target protein hamper the expression limiting the application of this tool. The aim of this study was finding the appropriate conditions for the expression of reluctant proteins that is the human neutral amino acid transporters ASCT2 and B0AT1, that have great relevance to human health in cancer therapy and in COVID-19 research, respectively. The cDNAs coding for the proteins of interest were cloned in the pCOLD I vector and different E. coli strains (BL21 codon plus RIL, and RosettaGami2) were cultured in absence or in presence of glucose (0.5-1%), at low temperature (15 degrees C), and low inducer concentrations (10-100 microM). Cell growth and protein production were monitored by optical density measurements and western blotting assay, respectively. Even though in different conditions, the expression of both amino acid transporters was obtained.Reducing the growth rate of specific E. coli strains by lowering the temperature and the IPTG concentration, together with the addition of glucose, two reluctant human neutral amino acid transporters have been expressed in E. coli. The results have a potentially great interest in drug discovery since ASCT2 is an acknowledged target of anticancer therapy, and B0AT1 together with ACE2 is part of a receptor for the SARS-Cov-2 RBD proteins. Twit Text FOAVip Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1) ????Mol Biol Rep http://www.kidney.de/pget.php?&tw=1&pmid=32772343 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32772343 #FOAvip English Wikipedia <ref>{{Cite pmid|32772343}}</ref> Low temperature bacterial expression of the neutral amino acid transporters SLC1A5 (ASCT2), and SLC6A19 (B0AT1) #MMPMID32772343 Galluccio M; Pantanella M; Giudice D; Brescia S; Indiveri C Mol Biol Rep 2020[Sep]; 47 (9): 7283-7289 PMID32772343show ga It is well established that Escherichia coli represents a powerful tool for the over-expression of human proteins for structure/function studies. In many cases, such as for membrane transporters, the bacterial toxicity or the aggregation of the target protein hamper the expression limiting the application of this tool. The aim of this study was finding the appropriate conditions for the expression of reluctant proteins that is the human neutral amino acid transporters ASCT2 and B0AT1, that have great relevance to human health in cancer therapy and in COVID-19 research, respectively. The cDNAs coding for the proteins of interest were cloned in the pCOLD I vector and different E. coli strains (BL21 codon plus RIL, and RosettaGami2) were cultured in absence or in presence of glucose (0.5-1%), at low temperature (15 degrees C), and low inducer concentrations (10-100 microM). Cell growth and protein production were monitored by optical density measurements and western blotting assay, respectively. Even though in different conditions, the expression of both amino acid transporters was obtained.Reducing the growth rate of specific E. coli strains by lowering the temperature and the IPTG concentration, together with the addition of glucose, two reluctant human neutral amino acid transporters have been expressed in E. coli. The results have a potentially great interest in drug discovery since ASCT2 is an acknowledged target of anticancer therapy, and B0AT1 together with ACE2 is part of a receptor for the SARS-Cov-2 RBD proteins. |Amino Acid Transport System ASC/genetics/*metabolism[MESH] |Amino Acid Transport Systems, Neutral/genetics/*metabolism[MESH] |Angiotensin-Converting Enzyme 2[MESH] |Betacoronavirus/*physiology[MESH] |COVID-19[MESH] |Cold Temperature[MESH] |Coronavirus Infections/*virology[MESH] |DNA, Complementary/genetics[MESH] |Drug Discovery[MESH] |Escherichia coli/genetics/*metabolism[MESH] |Gene Expression[MESH] |Humans[MESH] |Minor Histocompatibility Antigens/genetics/*metabolism[MESH] |Pandemics[MESH] |Peptidyl-Dipeptidase A/genetics/metabolism[MESH] |Pneumonia, Viral/*virology[MESH]Low temperature bacterial expression of the neutral amino acid transpo(epmc).pdf DeepDyve Pubget Overpricing