yeyiphi isantya se-centrifuge sokwahlula ii-ribosomes zasimahla
Isantya seCentrifuge sokwahlula iiRibosomes zasimahla: Indlela ebalulekileyo yoPhando lweBiology yeeMolekyuli
Intshayelelo:
Iiribosome zasimahla zidlala indima ebalulekileyo ekwakhiweni kweeproteni, kwaye ukuzihlukanisa nezinye izinto zeseli ngumsebenzi obalulekileyo kuphando lwebhayoloji yeemolekyuli. Eli nqaku liphonononga ukubaluleka kwesantya se-centrifuge ekwahlukaniseni ii-ribosome zasimahla kwaye liqaqambisa iinkqubo ezahlukeneyo kunye nezinto eziqwalaselwayo kule nkqubo.
I. Ukuqonda iiRibosomes zasimahla:
Ukuze kuqondwe ukubaluleka kwesantya se-centrifuge ekuhlukaniseni ii-ribosomes ezikhululekileyo, kubalulekile ukuqonda uhlobo kunye nemisebenzi yezi zinto zeseli. Ii-ribosomes ezikhululekileyo zizakhiwo ezincinci, ezinamaqhekeza afumaneka zidada ngokukhululekileyo kwi-cytoplasm, ezinoxanduva lokwenziwa kweeproteni ngaphakathi kwiseli. Azinalo ulwakhiwo olubotshelelwe yi-membrane, oluzahlula kwii-ribosomes ezinamathele kwi-endoplasmic reticulum. Ukuhlukaniswa kwee-ribosomes ezikhululekileyo kuvumela ukuqonda okunzulu ngeendima zazo ezibalulekileyo ekuveliseni iiproteni zeseli.
II. Imigaqo yeCentrifugation:
I-Centrifugation yindlela esetyenziswa kakhulu ukwahlula izinto ezahlukeneyo zeseli ngokusekelwe kubukhulu bazo, uxinano, kunye nobunzima beemolekyuli. Ibandakanya ukusetyenziswa kwamandla e-centrifugal, aveliswa ziisampuli ezijikelezayo ngesantya esiphezulu, ukwahlula inkunkuma yeseli, ii-organelles, kunye nee-macromolecules. Ukufezekisa ukwahlulwa kwe-ribosomes zasimahla, kubalulekile ukuphucula isantya kunye nobude be-centrifuge, njengoko ezi zinto zichaphazela kakhulu ukusebenza kakuhle kokwahlulwa.
III. Ukuphuculwa kwesantya seCentrifuge:
A. Ukuhlola iiRange zesantya:
Ukuchonga isantya esifanelekileyo se-centrifuge sokwahlula ii-ribosomes zasimahla kuxhomekeke kwizinto ezininzi, kubandakanya uhlobo lwesampulu kunye nenqanaba lobumsulwa elifunekayo. Ngokubanzi, abaphandi basebenza ngaphakathi koluhlu olubanzi lwesantya, beqala kwisantya esiphantsi ukuya ekususeni inkunkuma enkulu yeseli kwaye banda kancinci kancinci. Uluhlu lwesantya olufanelekileyo luhlala luphakathi kwe-10,000 ukuya kwi-100,000 xg, kunye notshintsho olusekelwe kwiimpawu ezithile zesampulu.
B. Iimpawu zeSampulu:
Iiseli kunye nezicubu ezahlukeneyo zinemilinganiselo eyahlukeneyo yobuninzi be-ribosome. Umzekelo, izicubu ezinesantya esiphezulu se-metabolism, njengeeseli zemisipha okanye zesibindi, zihlala ziqulathe ii-ribosomes ezininzi. Isantya se-centrifuge kufuneka sihlengahlengiswe ngokufanelekileyo ukuze kufunyanwe ukusebenza kakuhle kokwahlukana okufunekayo.
IV. IiProtocols zeCentrifugation zoKwahlukaniswa kweRibosome yasimahla:
A. Ukulungiselela iSampulu:
Ukuze kuphunyezwe ngempumelelo ii-ribosomes zasimahla, ukulungiselela isampuli ngokufanelekileyo kubalulekile. Iindlela zokucoca iiseli, ezinje nge-sonication okanye ukuphazamiseka kweeseli kusetyenziswa izicoci, kufuneka zisetyenziswe ukuqhekeza iiseli nokukhupha umxholo wazo. Emva koko, iisampuli ezicociweyo zidlula kuthotho lwamanyathelo okucoca ukuze kususwe ubumdaka, nto leyo evelisa i-supernatant ecacileyo equlethe ii-ribosomes.
B. Ukwahlukanisa iCentrifugation:
I-differential centrifugation, indlela esetyenziswa kakhulu, ibandakanya ukujikeleza okulandelelanayo okuliqela ngokonyusa isantya se-centrifuge. Le ndlela ivumela abaphandi ukuba bahlukanise ii-ribosomes kwezinye izinto zeseli ngokufanelekileyo. Ukujikeleza kokuqala okunesantya esiphantsi kususa ukungcola kweseli, kulandele isantya esiphakathi sokuhlukanisa ii-organelles kunye neeproteni ezinkulu zeseli. Okokugqibela, ukujikeleza okunesantya esiphezulu kusetyenziswa ukufumana iqhekeza elifunekayo elityebiswe kwii-ribosomes zasimahla.
C. I-Gradient Centrifugation:
I-gradient centrifugation isebenzisa i-density gradient media, efana ne-sucrose okanye i-cesium chloride, ukwahlula ii-ribosomes ngokusekelwe kubuninzi bazo obuthambileyo. Ngokubeka isampuli kwi-density gradient kunye ne-centrifuging ngesantya esilawulwayo, ii-ribosomes zinokwahlulwa ngesisombululo esongeziweyo. Le ndlela inika ubumsulwa obuphuculweyo kwaye ivumela ukwahlulwa kwee-subclasses ezahlukeneyo ze-ribosomes ngokusekelwe kumazinga azo okutsalwa.
V. Izinto Ezichaphazela Ukwenziwa Kakuhle kweSisantya seCentrifuge:
A. Umqulu weSampulu:
Umthamo wesampulu uchaphazela kakhulu ukwenziwa ngcono kwesantya se-centrifuge. Umthamo omkhulu wesampulu unokufuna amaxesha amade okufakwa kwe-centrifuge okanye isantya esiphezulu ukuqinisekisa ukwahlukana okwaneleyo. Nangona kunjalo, kubalulekile ukuqaphela ukuba isantya esigqithisileyo okanye amaxesha okufakwa kwe-centrifuge ahlala ixesha elide anokukhokelela kumonakalo we-ribosome kwaye achaphazele iimvavanyo ezilandelayo.
B. Uhlobo lweRotor:
Iirotha ze-centrifuge ezahlukeneyo zinemida yesantya eyahlukileyo kunye namandla aphezulu e-rotha acetyiswayo. Ukukhetha i-rotha efanelekileyo kubalulekile ukuze kuthintelwe ukonakala kwezixhobo kunye nokugcina ulwahlulo oluchanekileyo. Kubalulekile ukujonga incwadi yomsebenzisi ye-centrifuge kunye neengcebiso zomenzi xa ukhetha uhlobo lwe-rotha ngokusekelwe kwisantya esifunekayo kunye nomthamo wesampuli.
Isiphelo:
Ukuphucula isantya se-centrifuge kubalulekile ekuphuhlisweni ngempumelelo kwee-ribosomes zasimahla, nto leyo evumela ukufundwa kwendima yazo ekuhlanganisweni kweeproteni zeseli. Ngokulandela imigaqo ethile kunye nokuqwalasela iimpawu zesampulu, abaphandi banokuzahlula ngempumelelo ii-ribosomes zasimahla, bebonelela ngengqiqo ebalulekileyo ngemisebenzi yazo kunye nokusebenzisana kwazo kwindawo yeseli.
.