indlela yokwahlula i-chloroplast kunye ne-mitochondria nge-centrifugation
I-Centrifugation: Indlela Enamandla Yokwahlula i-Chloroplast kunye ne-Mitochondria
Intshayelelo:
I-Centrifugation yindlela esetyenziswa kakhulu kwi-molecular biology nakwi-biochemistry evumela ukwahlulahlula iinxalenye ezahlukeneyo zeseli ngokusekelwe kubukhulu bazo, imilo, kunye noxinano. Eli nqaku lijonga ukusetyenziswa kwe-centrifugation ukwahlula ii-chloroplast kunye ne-mitochondria, ii-organelles ezimbini ezibalulekileyo ezifumaneka kwiiseli ze-eukaryotic ezibandakanyeka kwimveliso yamandla kunye nezinye iinkqubo ezibalulekileyo zeseli.
Indima yeChloroplasts kunye neMitochondria kwiinkqubo zeSeli
Ii-chloroplast, ezikhoyo kwiiseli zezityalo, zinoxanduva lwe-photosynthesis, inkqubo eguqula amandla okukhanya abe ngamandla eekhemikhali agcinwe ngohlobo lwe-glucose. Le nkqubo ibalulekileyo yenza kube lula ukuveliswa kweoksijini kwaye ibalulekile ekuxhaseni ubomi eMhlabeni. I-Mitochondria, efumaneka kwiiseli zezityalo nezilwanyana, idla ngokubizwa ngokuba yindlu yamandla eseli. Zinoxanduva lokuphefumla kweeseli, ziguqula i-glucose ibe ngamandla asebenzisekayo kwiseli ngohlobo lwe-adenosine triphosphate (ATP).
Imfuneko Yokwahlukanisa iiChloroplast kunye neeMitochondria
Ukuqonda imisebenzi kunye neempawu ze-chloroplasts kunye ne-mitochondria kubalulekile kubaphandi abajonge ukufumana ulwazi kwiinkqubo zeseli kunye nokuphuhlisa unyango olunokwenzeka lwezifo ezahlukeneyo. Ukwahlula ezi organelles kumxube ontsonkothileyo wezinto zeseli kubalulekile ukuze kufunyanwe iisampulu ezicocekileyo zohlalutyo olongezelelweyo kunye novavanyo. Olu hluko lunokwenzeka nge-centrifugation.
Iziseko zeCentrifugation
Ukudibanisa i-centrifugation kubandakanya ukusetyenziswa kwamandla e-centrifugation ukwahlula izinto ngaphakathi kwesampuli. Isampuli ifakwa kwi-centrifuge, umatshini one-rotor ejikeleza ngesantya esiphezulu. Ngexesha lokujikeleza, amandla e-centrifugal abangela ukuba izinto ezixineneyo zibe yinkunkuma ezantsi, zenze i-pellet, ngelixa izinto ezingaxinenenga kangako zihlala kwi-supernatant. Isantya kunye nobude be-centrifugation, kunye nokukhethwa kweetyhubhu ze-centrifuge, ziyahluka ngokuxhomekeke kwisicelo esithile kunye nokwakheka kwesampuli.
I-Centrifugation eyahlukileyo yokwahlulwa kwe-Organelle
Eyona ndlela isetyenziswa kakhulu yokwahlula i-organelle yi-differential centrifugation. Le ndlela isekelwe kumgaqo wokwahlula isampuli ngokuyifaka kuthotho lwe-differential centrifugations ngesantya esonyukayo kunye nobude bexesha. Ngokulungisa iiparameter ze-centrifugation, abaphandi banokuzahlula ngempumelelo ii-chloroplast kunye ne-mitochondria kwezinye izinto zeseli.
Isikhokelo seNyathelo ngeNyathelo soKwahlula iiChloroplast kunye neeMitochondria
Inyathelo 1: Ukudibanisa
Inyathelo lokuqala kwi-differential centrifugation kukudibanisa isampuli ngendlela efanayo. Le nkqubo ibandakanya ukuqhekeza iiseli kancinci nokukhulula umxholo wazo, ukuvumela ukufikelela kwi-organelles. Iindlela ezahlukeneyo zingasetyenziswa ekuphazamiseni iiseli, njengokuphazamiseka koomatshini kunye nonyango lwe-nitrogen engamanzi.
Inyathelo lesi-2: I-Centrifugation yokwahluka
Emva kokuhlanganiswa kwe-homogenization, isampuli ifakwa kuthotho lwee-differential centrifugations. I-centrifugation yokuqala yesantya esiphantsi (1000-2000 xg) isusa inkunkuma enkulu yeseli kunye neeseli ezingaphukanga, ishiya ngasemva i-mitochondria eluhlaza kunye ne-chloroplast fraction. Le fraction ifakwa kwi-centrifugation yesantya esiphezulu (10,000-15,000 xg), nto leyo ekhokelela ekwahlulweni kwee-chloroplasts kwi-mitochondria.
Inyathelo lesi-3: Ukuphucula ubumsulwa
Ukuze kucocwe ngakumbi ii-chloroplast kunye nee-mitochondria fractions ezifunyenweyo, kunokwenziwa amanyathelo ongezelelweyo okucoca. Oku kuquka ukuphinda kuxhonywe i-pellet efunyenweyo emva kokucoca nganye kwi-buffer efanelekileyo kwaye kuphindwe inkqubo ngesantya esongezelelweyo kunye nobude bayo. Ezi centrifugations zongezelelweyo ziya kunceda ukususa naziphi na izinto ezingcolisayo eziseleyo kwaye zonyuse ubumsulwa bee-organelle fractions zokugqibela.
Isiphelo:
I-Centrifugation yindlela eguquguqukayo eye yaguqula icandelo lebhayoloji yeseli kwaye ibe sisixhobo esibalulekileyo sokwahlulahlula iinxalenye zeseli. Ngokusebenzisa i-differential centrifugation, abaphandi banokuhlulahlula ngempumelelo ii-chloroplast kunye ne-mitochondria, bebonelela ngeesampuli ezicocekileyo ezixabisekileyo zohlalutyo olongezelelweyo kunye novavanyo. Ukukwazi ukwahlula ezi organelles kuvula iingcango zokufunda imisebenzi yazo ethile kunye neendlela, okufaka isandla ekuqondeni ngcono iinkqubo zeseli kunye nokukhuthaza ukuqhubela phambili kwiinkalo ezahlukeneyo zesayensi.
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