Mokhoa oa ho fumana mahloriso a nepahetseng ka ho fetisisa?
Bakeng sa inducer IPTG (isopropyl-beta-d-thiogalactoside), ha mahloriso a le holimo, ho molemo. Mahloriso a nepahetseng a itšetlehile ka maemo a itseng a liteko le phello e lakatsehang ea induction.
Ka kakaretso, mahloriso a IPTG a sebediswa ho tloha ho 0.1-1 mM. Mahlakore a tlase a ka fokotsa ditlamorao tse mpe kgolong ya sele mme a ka fokotsa chefo e bakwang ke ho eketseha ha diprotheine tse rerilweng. Mahlakore a phahameng a ka baka moroalo o feteletseng wa metabolism ya sele, a ama kgolo ya sele le bokgoni ba ho hlahisa.
Mokhoa oa ho fumana hore na ho na le mahloriso a nepahetseng e ka ba ho lekola boemo ba tlhahiso ea protheine e reretsoeng ka ho etsa liteko tsa ho kenya IPTG maemong a fapaneng. Liteko tsa temo tse nyane li ka etsoa ho sebelisoa mefuta e fapaneng ea mahloriso a IPTG (mohlala, 0.1 mM, 0.5 mM, 1 mM, jj.) 'me phello ea tlhahiso maemong a fapaneng e ka lekoa ka ho lemoha boemo ba tlhahiso ea protheine e reretsoeng (mohlala, Western blot kapa fluorescence detection). Ho ea ka liphetho tsa liteko, mahloriso a nang le phello e ntle ka ho fetisisa ea tlhahiso a khethiloe e le mahloriso a nepahetseng ka ho fetisisa.
Ho phaella moo, o ka boela wa sheba dingolwa tse amehang kapa phihlelo ya dilaboratori tse ding ho utlwisisa mefuta e sebediswang haholo ya mahloriso a IPTG tlasa maemo a tshwanang a teko, ebe o ntlafatsa le ho fetola ho ya ka ditlhoko tsa teko.
Ho bohlokoa ho hlokomela hore mahloriso a nepahetseng a ka fapana ho latela litsamaiso tse fapaneng tsa tlhahiso, liprotheine tse reriloeng, le maemo a liteko, kahoo ho molemo ho ntlafatsa ho latela nyeoe ka 'ngoe.
Nako ea poso: Loetse-28-2023