Nzira yekuziva sei iyo yakakwana yekuisa pfungwa?
Kune iyo inducer IPTG (isopropyl-beta-d-thiogalactoside), iyo yakakwira iyo yekumisikidza, zviri nani.Iyo yakanyanya kusungirirwa inoenderana neiyo chaiyo yekuyedza mamiriro uye inodiwa induction maitiro.
Kazhinji, kuwanda kweIPTG kunoshandiswa muhuwandu hwe0.1-1 mM.Kudzika kwakadzika kunogona kuderedza mhedzisiro yakashata pakukura kwesero uye kunogona kuderedza cytotoxicity nekuda kwekuwedzeredza kwechinangwa cheprotein.Kukwirira kwakanyanya kunogona kukonzera kuwandisa kwesero metabolic mutoro, zvichikanganisa kukura kwesero uye kugona kutaura.
Iyo nzira yekuona iyo yakakwana yekuisa inogona kuve yekuongorora iyo yekutaura nhanho yeinonangwa puroteni nekuita IPTG induction bvunzo pane dzakasiyana siyana.Zviyedzo zvetsika diki-diki zvinogona kuitwa pachishandiswa huwandu hwe IPTG (eg 0.1 mM, 0.5 mM, 1 mM, zvichingodaro) uye mhedzisiro yekutaura pane zvakasiyana-siyana inogona kuongororwa nekuona danho rekutaura reprotein yakanangwa (semuenzaniso Western. blot kana fluorescence kuonekwa).Zvinoenderana nemhedzisiro yekuyedza, iyo yekumisikidza ine yakanakisa kutaura mhedzisiro yakasarudzwa seyakanyanya kugadzirisa.
Pamusoro pezvo, iwe unogona zvakare kutarisa kune akakodzera mabhuku kana ruzivo rwemamwe marabhoritari kuti unzwisise inowanzoshandiswa IPTG yekumisikidza renji pasi pemamiriro ekuyedza akafanana, wobva wawedzera uye kugadzirisa zvinoenderana nezvinodiwa zvekuedza.
Izvo zvakakosha kuti uzive kuti iyo optimal concentration inogona kusiyana zvichienderana neakasiyana ekutaura masisitimu, anonangwa mapuroteni, uye kuyedza mamiriro, saka zviri nani kukwirisa pane imwe-ne-kesi hwaro.
Nguva yekutumira: Sep-28-2023