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大香蕉综合伊人91新人

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Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA

JoVE Journal
Biochemistry

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JoVE Journal Biochemistry

Single-Molecule Imaging of EWS-FLI1 Condensates Assembling on DNA

??????? ????? ?????? ?????? ?????? EWS-FLI1 ??? ????? ??????

Full Text

2,505 Views

07:05 min

September 8, 2021

DOI: 10.3791/62974-v

Linyu Zuo*¹, Jiawei Ding*¹, Zhi Qi¹

¹Center for Quantitative Biology, Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies,Peking University

Please note that some of the translations on this page are AI generated. Click here for the English version.

Summary

??? ? ??? ??????? ????? ??????? ?????? ?????? ? ????? ????? ?????? ? ?????? ?????? ?????????? ??????? ??????? EWS-FLI1 ???????? ??? ????? ??????.

Transcript

??? ???? ?????? ???? ???? ??????? ????? ?????? ?????? ????? ??? ????? ?????? ???? ??? ??????? ?????? ??????? ??????????? ?????? ????? ?? ????? ?????? ???? ??? ??????? ????? ????? ??? ????? ??????. EWS-FL1 ?? ??? ????? ???????? ?????? ?????? ?? ????? ??? ??????? ??????. ???? ?? ???? ????? ??????? ??? ??? ??? ????? ?????? ?????? ?????? EWS-FL1 ???????? ???? ??????? ??????? ?? ??????? ????????? ? ????? ???? ????? ???????.

???? ????? ??? ??????? ?????? ?? ?????? ??? ?? ?????? ?? ??????? ? ??? P53 ? MED1. ?????? ???? ???? ?????? ???? ?????? ????? ?????? ????? ????? ?????? ? ?? ?????? ?????? ??????? ???????? ?? ??????? ??????. ?? ???? ???? ?????? ? 2.5 ??????? ?? ?????? ?????? ?????? ???????? ?????? ??? ????? ??????? ? ??? ???? ??? ???? ????? ?? ???? ??????.

???? ??????? ?? ??????? ???? ??????? ???? ?? ????? ???????? ??????? ???? ???? ?? ????? ???? ??? ????? ??? ?? ????. ?????? ? ??? ??? ???? ?????? ? 2.5 ?? ?? ?????? ?????? ?????? ???? ? ????????? ?? ???? ????? ?????? ???? 30 ?????. ????? ??????? ? ?? ?????? ???? BSA ?????? ??? ?? ????? ?? ?????? ????.

?? ??? 30 ????? ?? ?????? ? ???? ???? ?????? ? 2.5 ??????? ?? ?????? ?????? BSA ?? ???? ???????. ??? ??? ? ?? ???? 800 ???????? ?? ???? ????????????? ?? ???? ?????? ?? ???? ??????? ?? ?????? ?? ????? ???? 10 ????? ??? ?? ????. ?? ???? ???? ?????? ? 2.5 ??????? ?? ????? BSA ?????? ???? ?????? ????????????? ?????.

??? ??? ? ?? ?????? ????? ?????? ???????? Lambda ?????? ??????? ???????? ???? BSA ?????? ????? ???? ???? ???? ?? ??? ?????. ????? ????? ? ?? ?????? ?????? ?? ???? ????? ???????? ????? ??????? ?????? ???????. ?? ???? ??????? ? 10 ??????? ?? ????? ?????? ??????? ????? ??????? ????? ??????? ?????? ????? ????? ???? 2? ? 99? ???????.

??? ??? ? ???? ?? ?? ??? ?? 20 ??????? ?? ?????? ?????? ??????? ?? ???? ??? 30 ???????. ?? ?????? ???? ?????? ?? ????? ?????? ??? ???????? ????? ??????? ???????. ???????? ???? ???? 0.03 ??????? ?? ??????? ? ???? ?????? ????? ?????? ??? ?????? ???? 10 ?????.

?? ?? ?????? ????? ???? ??????? ??????? ???????? ???? 30 ?????. ?? ???? ?????? ? ??? ???? ????? ?????? ????????? ???? ?????. ?????? ????? ????? EWS-FLI1 ??? ????? ????? ?????? ? ???? ?????? ??????? ????? ?? ????? ??????? ???????? ?????? ??? ?????? ?????? ??? ????????.

?? ?? ?????? ?????? ????? 0.2 ??????? ?? ??????? ?????? ????? ?????? ????? ????? ?????? ???????? ???? ????? ??? ????? ???? 10 ?????. ??? ??? ? ?? ?????? ?????? mCherry EWS-FLI1 ???????? ???? ??????? ?????? ?????? 100 ??????? ?? 100 ????????. ?? ?? ?????? ???? ???????? ?? ???? ?????? ???????? ???? ?????? ??? 100 ???????? ??? ?????? ???? ?????? ??? 0.4 ??????? ?? ???????.

??? ????? ?????? 488 ??????? ? ?? ?????? ??? ???? ??? ????? ?????? ?? ???? ????? ????? ?????? ?????? ??????? ???? ????? ???? ?????? ????? ?????? ????????. ?? ???? ???? ?????? ??? 10? ????? 488 ??????? ? 20? ????? 561 ???????. ????????? ????? ?????? ? ?? ????? ??? ?????? ???????? ?????? ?? ???????.

???? ?? ?????? ??? ????? ??? ?????? ??????? ???????? ?? ?? ???? 488 ? 561 ??????? ?? ??? ????. ?? ?? ?????? ?????? ?? ????? ?????? ??? ??? ????? ?????? ?????? ?????? ??????? ???? ???? ???????? ?? ???? ?????? ??? 60 ?????. ?????? EWS-FLI1 ??????? ? ????? ?? ??? ???? ?????? ???????? ???? ??????? ?????? ???? ??? ????? ?? ????? ?????? 561 ??????? ???.

?? ???? ?????? ???????? ??? 37 ???? ????? ???? 10 ?????. ??? 10 ????? ? ?? ?????? ?????? ????? 0.4 ??????? ?? ??????? ?????? ????? ?????? ??????? ??????? ??? ??? ??? ?????? ??????? ??? ?????? ?????? ?????? ??? ?????? ??????? ????? ?????? ?????? EWS-FLI1. ?? ???? ?????? EWS-FLI1 ?? ???? ????? ?? ?????? mCherry ??????? EWS-FLI1 ???? ?? ?????? ????? ???????? ???? 561 ???????.

???? ???? ??????? ???????? ??????? EWS-FLI1 ?? ???? 25 ?????? GGAA ?? ????? ????? ?????? ??????. ?? ????? ?????? mCherry EWS-FLI1 ????????? ?? ????? ????? ?????? ?? ???? ?????? ????? ?????? ?????????? ???????? ???? ????? ?????? ?? ????? ????? 25 GGAA ???? ?????. ????? ??? ?????? ????? EWS-FLI1 ?? 20 ??? 500 ??????? ? ???? ??? EWS-FLI1 ???? ????.

?? ??? ?? ??????? ?? ??? FLI ???? DBD ??? ????? ????? ???? ?????????? ????? ?????? ??????? GGAA ? ??? ???? ??? ?? EWS-FLI1 ???? ?????? ??? ????? ??????. ??? ?? ???? ????? ????? ??? ?????? ??? ????? ????????? ?????? ?????? ?? ??????? ???????? ??? ???? ??????. ?? ????? ????? MSA ?????? ?? ?? ???? ????? ?????? ????? ??? ??? ??????? ???????? ???????? ?? ???????.

???? ??? ??????? ??????? ???????? ?? ??? ???? ???????? ????? ????? ???????? ?? ????? ????? ???????? ??? ?????.

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