Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural dif...Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.展开更多
A 551-bp hIL-11 gene fragment that includes no nucleotide sequences encoding signalpolypeptide and the initial 8 amino acids of the mature protein was cloned into a high-level expression vectorpEx31B of E.coli.The aut...A 551-bp hIL-11 gene fragment that includes no nucleotide sequences encoding signalpolypeptide and the initial 8 amino acids of the mature protein was cloned into a high-level expression vectorpEx31B of E.coli.The authors identified the recombinant plasmid,designated pEx31-IL11,by restriction endonu-cleases digestion and DNA sequencing.The resulting recombinant plasmids were then used to transform E.colistrain HB101,and expression in the PL promoter system,which is temperature-regulated,was achieved.The ex-pressed fusion protein amounts to 50% of total bacterial proteins.The hIL-11 protein expressed in E.coliwas fused to the N-terminal 99 amino acids of the MS2 polymerase to form the inclusion body.Theserecombinant proteins can be purified to about 80% by extracting inclusion body with urea.OneIL-6-dependent cell line 7 TD1 was used for bioassay.The recombinant hIL-11 protein was preliminarily puri-fied and renatured to a specific activity of 10~5U/mg,even in the presence of an excess of a neutralizing an-ti-IL-6 antibody.展开更多
基金supported by the National Natural Science Foundation of China,No.31340024
文摘Overexpression of receptor-interacting protein 140（RIP140） promotes neuronal differentiation of N2 a cells via extracellular regulated kinase 1/2（ERK1/2） signaling.However,involvement of RIP140 in human neural differentiation remains unclear.We found both RIP140 and ERK1/2 expression increased during neural differentiation of H1 human embryonic stem cells.Moreover,RIP140 negatively correlated with stem cell markers Oct4 and Sox2 during early stages of neural differentiation,and positively correlated with the neural stem cell marker Nestin during later stages.Thus,ERK1/2 signaling may provide the molecular mechanism by which RIP140 takes part in neural differentiation to eventually affect the number of neurons produced.
文摘A 551-bp hIL-11 gene fragment that includes no nucleotide sequences encoding signalpolypeptide and the initial 8 amino acids of the mature protein was cloned into a high-level expression vectorpEx31B of E.coli.The authors identified the recombinant plasmid,designated pEx31-IL11,by restriction endonu-cleases digestion and DNA sequencing.The resulting recombinant plasmids were then used to transform E.colistrain HB101,and expression in the PL promoter system,which is temperature-regulated,was achieved.The ex-pressed fusion protein amounts to 50% of total bacterial proteins.The hIL-11 protein expressed in E.coliwas fused to the N-terminal 99 amino acids of the MS2 polymerase to form the inclusion body.Theserecombinant proteins can be purified to about 80% by extracting inclusion body with urea.OneIL-6-dependent cell line 7 TD1 was used for bioassay.The recombinant hIL-11 protein was preliminarily puri-fied and renatured to a specific activity of 10~5U/mg,even in the presence of an excess of a neutralizing an-ti-IL-6 antibody.
