Incazelo yamagama ayisisekelo ebhayoloji yamangqamuzana
1. I-cDNA kanye ne-cccDNA: I-cDNA iyi-DNA enemicu ekabili ehlanganiswe yi-reverse transcriptase evela ku-mRNA;I-cccDNA iyi-plasmid eyindilinga eyi-DNA eyindilinga ephindwe kabili engena ku-chromosome.
2. Iyunithi yokugoqa evamile: iyunithi yesakhiwo sesibili seprotheni i-α-helix ne-β-sheet ingakha amabhulokhi esakhiwo ngamalungiselelo akhethekile wejometri ngokusebenzisa ama-polypeptide ahlukahlukene axhumayo.Lolu hlobo lokugoqa okunqunyiwe luvame ukubizwa ngokuthi isakhiwo sesibili esikhulu.Cishe zonke izakhiwo zemfundo ephakeme zingachazwa yilezi zinhlobo zokugoqa, ngisho nezinhlobo zazo ezihlanganisiwe, ngakho-ke zibizwa nangokuthi amayunithi okugoqa ajwayelekile.
3. CAP: cyclic adenosine monophosphate (cAMP) receptor protein CRP (cAMP receptor protein), inkimbinkimbi eyakhiwe ngemva kwenhlanganisela ye-cAMP ne-CRP ibizwa ngokuthi activating protein CAP (cAMP activated protein)
4. Ukulandelana kwe-Palindromic: Ukulandelana okuphambene okuphambene kwengxenye yesiqephu se-DNA, ngokuvamile isayithi le-enzyme evimbela.
5. I-micRNA: I-RNA ephazamisayo ehambisanayo noma i-RNA ye-antisense, ehambisana nokulandelana kwe-mRNA futhi engavimbela ukuhunyushwa kwe-mRNA.
6. I-Ribozyme: I-RNA enomsebenzi we-catalytic, edlala indima ye-autocatalytic kunqubo yokuhlanganisa i-RNA.
7. I-Motif: Kunezifunda ezithile zasendaweni ezinomumo ofanayo-ntathu kanye ne-topology esakhiweni sendawo samangqamuzana amaprotheni.
8. I-peptide yesignali: i-peptide enezinsalela ze-amino acid ezingu-15-36 ku-N-terminus ngesikhathi sokuhlanganiswa kwamaprotheni, eqondisa i-transmembrane yeprotheni.
9. I-Attenuator: Ukulandelana kwe-nucleotide phakathi kwesifunda somsebenzisi kanye nofuzo lwesakhiwo olunqamula ukuloba.
10. I-Magic Spot: Uma amagciwane ekhula futhi ehlangabezana nokuntuleka okuphelele kwama-amino acid, amabhaktheriya azokhiqiza impendulo ephuthumayo ukuze amise ukubonakaliswa kwazo zonke izakhi zofuzo.Amasignali akhiqiza le mpendulo ephuthumayo i-guanosine tetraphosphate (ppGpp) ne-guanosine pentaphosphate (pppGpp).Iqhaza le-PpGpp ne-pppGpp akuyona nje i-opereni eyodwa noma ezimbalwa, kodwa ithinta inani elikhulu lazo, ngakho-ke zibizwa ngokuthi ama-super-regulators noma izindawo zomlingo.
11. Isici somgqugquzeli okhuphuka nomfula: ibhekisela ekulandelaneni kwe-DNA edlala indima yokulawula emsebenzini womthuthukisi, njenge-TATA esifundeni -10, i-TGACA esifundeni -35, izithuthukisi, nezixhumi.
12. I-DNA probe: ingxenye enelebula ye-DNA enokulandelana okwaziwayo, esetshenziswa kakhulu ukuthola ukulandelana okungaziwa kanye nezakhi zofuzo zesikrini.
13. Ukulandelana kwe-SD: Iwukulandelana okubophezelayo kwe-ribosome ne-mRNA, elawula ukuhumusha.
14. I-Monoclonal Antibody: I-antibody esebenza kuphela ngokumelene nesihlukanisi esisodwa se-antigen.
15. I-Cosmid: Ivetha ye-DNA yangaphandle eyakhiwe ngokwenziwa egcina izifunda ze-COS kuzo zombili iziphetho zephaji futhi ixhunywe ku-plasmid.
16. Ukuhlolwa kwendawo okumhlophe okwesibhakabhaka: Isakhi sofuzo se-LacZ (i-encoding β-galactosidase), i-enzyme ingabola i-chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) ukuze ikhiqize okuluhlaza okwesibhakabhaka, ngaleyo ndlela yenze uhlobo lube luhlaza okwesibhakabhaka.Lapho kufakwa i-DNA yangaphandle, isakhi sofuzo se-LacZ asikwazi ukuvezwa, futhi uhlobo lumhlophe, ukuze kuhlolwe amagciwane aphinde ahlanganiswe.Lokhu kubizwa nge-blue-white screening.
17. I-Cis-acting element: Ukulandelana okukhethekile kwezisekelo ku-DNA ezilawula ukusho kofuzo.
18. I-Klenow enzyme: Ingxenye enkulu ye-DNA polymerase I, ngaphandle kokuthi umsebenzi we-5' 3' we-exonuclease ukhishwa ku-DNA polymerase I holoenzyme.
19. I-PCR eqinile: isetshenziselwa ukukhulisa i-DNA yokuthakaselayo ngokulandelana okwaziwayo ekugcineni.Umsila we-poly-dG wengezwe ekugcineni kokulandelana okungaziwa, kwase kusetshenziswa i-poly-dC nokulandelana okwaziwayo njengeziqalo zokukhulisa i-PCR.
20. Iphrotheni ye-Fusion: Isakhi sofuzo seprotheni ye-eukaryotic sixhunywe nesakhi sofuzo sangaphandle, futhi iphrotheni eyakhiwe ukuhunyushwa kweprotheni yofuzo lwangempela kanye namaprotheni angaphandle ivezwa ngesikhathi esifanayo.
Eminye imigomo yebhayoloji yamangqamuzana
1. Imephu ebonakalayo ye-DNA iwuhlelo lapho izingcezu ze-molecule ye-DNA zihlelwa khona.
2. Ukuqhekeka kwe-RNase kuhlukaniswe izinhlobo ezimbili (i-autocatalysis) kanye (ne-heterocatalysis).
3. Kunezici ezintathu zokuqalisa ku-prokaryotes yilezi (IF-1), (IF-2) kanye (IF-3).
4. Amaprotheni e-Transmembrane adinga isiqondiso (ama-peptide esignali), futhi indima yama-protein chaperones (isiza ukugoqa iketango le-peptide lingene ekwakhiweni komdabu kwephrotheni).
5. Ama-elementi kubathuthukisi ngokuvamile angahlukaniswa abe izinhlobo ezimbili: (ama-elementi ayinhloko womphromotha) kanye (nama-elementi womphromotha okhuphuka nomfula).
6. Okuqukethwe kocwaningo lwebhayoloji yamangqamuzana ikakhulukazi kufaka izingxenye ezintathu: (i-structural molecular biology), (i-gene expression and regulation), kanye (nobuchwepheshe bokuhlanganiswa kabusha kwe-DNA).
7. Ukuhlola okubili okubalulekile okubonisa ukuthi i-DNA iyimpahla yofuzo yilezi (i-pneumococcus infection yamagundane) kanye (i-T2 phage infection ye-Escherichia coli).okungenzeka).
8. Kukhona umehluko omkhulu phakathi kwe-hnRNA ne-mRNA: (i-hnRNA ihlukaniswe ngenqubo yokuguqulelwa ku-mRNA), (ukuphela okungu-5' kwe-mRNA kwengezwa ngekepisi ye-m7pGppp, futhi kukhona i-polyadenylation eyengeziwe ekugcineni kuka-3' we-mRNA acid (polyA) umsila).
9. Izinzuzo zendlela yama-multi-subunit amaprotheni yilezi (i-subunit iyindlela yokonga yokusetshenziswa kwe-DNA), (inganciphisa umthelela wamaphutha angahleliwe ekuhlanganiseni kwamaprotheni emsebenzini wamaprotheni), (umsebenzi ungaba ngempumelelo kakhulu futhi ngokushesha avulwe futhi avalwe).
10. Okuqukethwe okuyinhloko kwendlela yokugoqa amaprotheni ithiyori yokuqala ye-nucleation ihlanganisa (i-nucleation), (ukucebisa kwesakhiwo), (ukuhlelwa kabusha kokugcina).
11. I-galactose inomphumela okabili kubhaktheriya;ngakolunye uhlangothi (ingasetshenziswa njengomthombo wekhabhoni wokukhula kwamaseli);ngakolunye uhlangothi (kubuye kube ingxenye yodonga lwamaseli).Ngakho-ke, umgqugquzeli ozimele we-cAMP-CRP i-S2 uyadingeka ukuze kuhlanganiswe unomphela ezingeni elingemuva;ngesikhathi esifanayo, umgqugquzeli oncike ku-cAMP-CRP i-S1 uyadingeka ukuze ulawule ukuhlanganiswa kwezinga eliphezulu.Ukuloba kuqala ku-( S2 ) ngo-G futhi ukusuka ku- ( S1 ) ngaphandle kuka-G.
12. Ubuchwepheshe be-Recombinant DNA bubuye baziwe ngokuthi (i-gene cloning) noma (i-molecular cloning).Umgomo omkhulu uwu (ukudlulisa ulwazi lofuzo lwe-DNA kwenye into ephilayo luluyise kwenye into ephilayo).Ukuhlolwa okujwayelekile kokuhlanganiswa kabusha kwe-DNA kuvame ukuhlanganisa lezi zinyathelo ezilandelayo: (1) Khipha isakhi sofuzo esiqondiwe (noma isakhi sofuzo sangaphandle) somzimba onikelayo, bese usixhuma nge-enzyme kwenye i-molecule ye-DNA (i-cloning vector) ukuze wakhe i-molecule entsha ye-DNA.② I-molecule ye-DNA ehlanganisiwe idluliselwa kuseli lomamukeli futhi iphindaphindwe kuseli lomamukeli.Le nqubo ibizwa ngokuthi ukuguqulwa.③ Khipha futhi ukhombe lawo maseli abamukeli abamunce i-DNA recombinant.④ Hlakulela amaseli aqukethe i-DNA ehlanganisiwe ngobuningi ukuze uthole ukuthi ufuzo losizo lwangaphandle luyavezwa yini.
13. Kunezinhlobo ezimbili zokuphindaphinda kwe-plasmid: lezo ezilawulwa ngokuqinile yi-host cell protein synthesis zibizwa ngokuthi (ama-plasmids aqinile), futhi lawo angalawulwa ngokuqinile yi-host cell protein synthesis abizwa ngokuthi (ama-plasmids akhululekile).
14. Uhlelo lwe-PCR lokusabela kufanele lube nezimo ezilandelayo: a.Iziqalo ze-DNA (cishe izisekelo ezingu-20) ezinokulandelana okuhambisanayo ekugcineni kwemigqa emibili yofuzo oluqondiwe okufanele luhlukaniswe.b.Ama-Enzymes anokuqina okushisayo njenge: TagDNA polymerase.c, dNTPd, DNA ukulandelana kokuthakaselayo njengesifanekiso
15. Inqubo eyisisekelo yokusabela ye-PCR ihlanganisa izigaba ezintathu: (i-denaturation), (i-annealing), kanye (nesandiso).
16. Inqubo eyisisekelo yezilwane ezishintshashintshayo ngokuvamile ihlanganisa: ①Ukwethulwa kofuzo lwangaphandle oluhlanganisiwe ku-nucleus yeqanda elivundisiwe noma i-embryonic stem cell;②Ukufakelwa kweqanda elivundisiwe elivundisiwe noma i-embryonic stem cell esibelethweni sowesifazane;③Ukuthuthuka nokukhula okuphelele kwe-embryonic Enzalweni enamafuzo angaphandle;④ Sebenzisa lezi zilwane ezingakhiqiza amaprotheni angaphandle njengemfuyo yokuzalanisa ukuzalanisa imigqa emisha ye-homozygous.
17. Imigqa yamaseli e-Hybridoma ikhiqizwa ngamaseli e-hybridizing (spleen B) anamaseli (i-myeloma), futhi njengoba (amangqamuzana e-spleen) angasebenzisa i-hypoxanthine futhi (amangqamuzana amathambo) ahlinzeka ngemisebenzi yokuhlukaniswa kwamaseli, angakhuliswa ku-HAT medium.khula.
18. Ngokujula kocwaningo, isizukulwane sokuqala samasosha omzimba sibizwa ngokuthi (ama-polyclonal antibodies), isizukulwane sesibili (amasosha omzimba angama-monoclonal), kanye nesizukulwane sesithathu (amasosha omzimba wobunjiniyela bofuzo).
19. Njengamanje, ubunjiniyela bezakhi zofuzo zamagciwane ezinambuzane bugxile kakhulu ku-baculovirus, ebonakaliswa ekusungulweni kwe- (exogenous toxin gene);(izakhi zofuzo eziphazamisa umjikelezo wokuphila ovamile wezinambuzane);(ukuguqulwa kwezakhi zofuzo zegciwane).
20. Izinto zamaprotheni ezishintshayo ezihambisana nezinto ezivamile i-TATA, GC, ne-CAAT kumgqugquzeli we-RNA polymerase II encelisayo yi-(TFIID), (SP-1) kanye (CTF/NF1), ngokulandelanayo.
amashumi amabili nanye.Izinto eziyisisekelo zokuloba ze-RNA polymerase Ⅱ yilezi, TFⅡ-A, TFⅡ-B, TFII-D, TFⅡ-E, futhi ukulandelana kwazo okubophayo kuthi: (D, A, B, E).Lapho umsebenzi we-TFII-D uwukuthi (ubophezela ebhokisini le-TATA).
amashumi amabili nambili.Iningi lezinto ezilotshiweyo ezibophezela ku-DNA zisebenza ngendlela yama-dimers.Izizinda ezisebenzayo zezinto ezilotshiweyo ezibophezela ku-DNA ngokuvamile yilezi ezilandelayo (i-helix-turn-helix), (i-zinc finger motif), (i-basic-leucine) i-zipper motif).
amashumi amabili nantathu.Kunezinhlobo ezintathu zezindlela zokunqanda i-endonuclease cleavage: (sika ku-5' side ye-symmetry axis ukuze ukhiqize iziphetho ezinamathelayo ezingu-5'), (sika ohlangothini olu-3' lwe-symmetry axis ukuze ukhiqize iziphetho ezinamathelayo ezingu-3' (sika ku-axis yokulinganisa ukuze ukhiqize amasegimenti ayisicaba) ).
amashumi amabili nane.I-Plasmid DNA inokucushwa okuthathu okuhlukene: (ukucushwa kwe-SC), (ukucushwa kwe-oc), (ukucushwa kwe-L).Eyokuqala ku-electrophoresis yi-(SC configuration).
25. Izinhlelo zokuvezwa kwezakhi zofuzo zangaphandle, ikakhulukazi (Escherichia coli), (Yeast), (Isinambuzane) kanye (nethebula lamaseli ezilwane ezincelisayo).
26. Izindlela ezisetshenziswa kakhulu ezilwaneni ezishintshashintshayo yilezi: (indlela yokutheleleka nge-retroviral), (indlela ye-DNA microinjection), (indlela ye-embryonic stem cell).
Isicelo sebhayoloji yamangqamuzana
1. Yisho imisebenzi yama-RNA angaphezu kwama-5?
Dlulisa i-RNA tRNA Dlulisa i-amino acid I-Ribosome RNA rRNA I-Ribosome yenza isithunywa i-RNA mRNA isifanekiso se-protein synthesis ye-Heterogeneous Nuclear RNA hnRNA Isandulela se-mRNA ekhulile yenukliya i-RNA snRNA Ihileleke ekuhlanganiseni kwe-hnRNA kanye ne-cytoplasmic RNA i-RNA ehlanganisiwe i-RNA i-Plasic RNA encane-RNA i-RNA ehlanganisiwe izingxenye zomzimba zokuqaphela isignali Antisense RNA anRNA/micRNA Ilawula ukusho kofuzo I-Ribozyme RNA I-RNA esebenza nge-enzyme
2. Uyini umehluko omkhulu phakathi kwabagqugquzeli be-prokaryotic kanye ne-eukaryotic?
I-Prokaryotic TTGACA --- TATAAT------Indawo Yokuqalisa-35 -10 I-Eukaryotic Enhancer---GC ---CAAT----TATAA-5mGpp-Indawo Yokuqalisa-110 -70 -25
3. Yiziphi izici eziyinhloko zokwakha ama-plasmids emvelo?
Ama-plasmid emvelo avame ukuba neziphambeko, ngakho-ke awafanele ukusetshenziswa njengabathwali bobunjiniyela bofuzo, futhi kufanele ashintshwe futhi akhiwe: a.Engeza izakhi zofuzo zomaka zokukhetha ezifanele, ezifana nambili noma ngaphezulu, okulula ukuzisebenzisa uma ukhetha, ngokuvamile izakhi zofuzo zama-antibiotic.b.Khulisa noma unciphise izindawo zokusika ama-enzyme ukuze kube lula ukuhlanganisa.c.Nciphisa ubude, unqume izingcezu ezingadingekile, uthuthukise ukusebenza kahle kokungenisa futhi ukhulise umthamo wokulayisha.d.Shintsha i-replicon, ukusuka kokuqinile ukuya kokuxekethile, ukusuka kumakhophi ambalwa ukuya kwamakhophi amaningi.e.Engeza izakhi zofuzo ezikhethekile ngokwezidingo ezikhethekile zobunjiniyela bofuzo
4. Nikeza isibonelo sendlela yokuhlolwa okuhlukile kwe-cDNA eqondene nezicubu ezithile?
Izinhlobo zamangqamuzana amabili ziyalungiswa, isakhi sofuzo esiqondiwe sivezwa noma sivezwe kakhulu kwelinye lamaseli, futhi isakhi sofuzo esiqondiwe asivezwa noma sivezwe kancane kwelinye iseli, bese kuthi isakhi sofuzo esihlosiwe sitholwa ngokuhlanganisa nokuqhathanisa.Isibonelo, phakathi nokuvela nokuthuthukiswa kwezimila, amaseli wesimila azokwethula ama-mRNA ngamaleveli enkulumo ahlukene kunamaseli avamile.Ngakho-ke, izakhi zofuzo ezihlobene nesimila zingahlolwa ngokuhlanganisa okuhlukile.Indlela yokungeniswa ingase isetshenziselwe ukuhluza izakhi zofuzo okukhulunywa ngakho kuye.
5. Isizukulwane kanye nokuhlolwa kwemigqa yeseli ye-hybridoma?
Amaseli we-spleen B + amaseli e-myeloma, engeza i-polyethylene glycol (PEG) ukuze akhuthaze ukuhlanganiswa kwamangqamuzana, kanye namaseli e-splenic B-myeloma fusion akhule ku-HAT medium (equkethe i-hypoxanthine, aminopterin, T) ayaqhubeka nokwandisa ukondla.I-cell fusion iqukethe: amaseli e-spleen-spleen fusion: awakwazi ukukhula, amangqamuzana e-spleen awakwazi ukukhuliswa nge-vitro.Amaseli okuhlanganisa amathambo: awakwazi ukusebenzisa i-hypoxanthine, kodwa angahlanganisa i-purine ngendlela yesibili esebenzisa i-folate reductase.I-Aminopterin inhibits folate reductase futhi ngaleyo ndlela ayikwazi ukukhula.Amaseli e-Bone-spleen fusion: angakhula ku-HAT, amaseli ubende angasebenzisa i-hypoxanthine, namaseli amathambo ahlinzeka ngokusebenza kokuhlukaniswa kwamaseli.
6. Uyini umgomo kanye nendlela yokunquma ukwakheka okuyinhloko kwe-DNA ngendlela ye-didioxy terminal termination (indlela ye-Sanger)?
Isimiso siwukusebenzisa i-nucleotide chain terminator—2,,3,-dideoxynucleotide ukuze kunqandwe ukunwetshwa kwe-DNA.Njengoba ingenayo i-3-OH edingekayo ukuze kwakhiwe ibhondi ye-3/5/phosphodiester, uma isifakwe ochungechungeni lwe-DNA, uchungechunge lwe-DNA alukwazi ukunwetshwa.Ngokomgomo wokumataniswa kwesisekelo, noma nini lapho i-DNA polymerase idinga i-dNMP ukuze ibambe iqhaza ochungechungeni lwe-DNA olwandisiwe, kunamathuba amabili, okukodwa ukubamba iqhaza ku-ddNTP, okuholela ekunqanyulweni kwesandiso seketango le-deoxynucleotide;enye iwukuba iqhaza ku-dNTP , ukuze uchungechunge lwe-DNA lusakwazi ukuqhubeka nokunwetshwa kuze kuhlanganiswe i-ddNTP elandelayo.Ngokwale ndlela, iqoqo lezingcezu ze-DNA ezinobude obuhlukene ezigcina nge-ddNTP zingatholakala.Indlela iwukuhlukanisa ngamaqembu amane ngokulandelana i-ddAMP, i-ddGMP, i-ddCMP, ne-ddTMP.Ngemuva kokusabela, i-polyacrylamide gel electrophoresis ingafunda ukulandelana kwe-DNA ngokuya ngamabhendi okubhukuda.
7. Uyini umphumela omuhle wokulawula we-activator protein (CAP) ekulotshweni?
Iphrotheni ye-Cyclic adenylate (cAMP) ye-receptor CRP (iphrotheni ye-cAMP ye-receptor), inkimbinkimbi eyakhiwe inhlanganisela ye-cAMP ne-CRP ibizwa ngokuthi i-CAP (i-cAMPactivated protein).Lapho i-E. coli ikhuliswa esimeni esintula i-glucose, ukuhlanganiswa kwe-CAP kuyanda, futhi i-CAP inomsebenzi wokuvuselela abagqugquzeli abafana ne-lactose (Lac).Abanye abathuthukisi abancike ku-CRP abanaso isici esijwayelekile sokulandelana kwesifunda -35 (TTGACA) abathuthukisi abavamile abanaso.Ngakho-ke, kunzima nge-RNA polymerase ukubopha kuyo.Ukuba khona kwe-CAP (umsebenzi): kungathuthukisa kakhulu ukubopha okungaguquki kwe-enzyme kanye nomgqugquzeli.Ikhombisa ngokuyinhloko izici ezimbili ezilandelayo: ① I-CAP isiza i-molecule ye-enzyme ukuthi ihambe ngendlela efanele ngokushintsha ukwakheka komgqugquzeli nokusebenzelana ne-enzyme, ukuze ihlangane nesifunda -10 futhi idlale indima yokufaka esikhundleni somsebenzi wesifunda -35.I-②CAP ingaphinda ivimbele ukuboshwa kwe-RNA polymerase kwamanye amasayithi ku-DNA, ngaleyo ndlela ikhulise amathuba okubophezela kumdidiyeli wayo othile.
8. Yiziphi izinyathelo ezivame ukufakwa ocwaningweni oluvamile lokuhlanganisa kabusha i-DNA?
a.Khipha isakhi sofuzo esiqondiwe (noma isakhi sofuzo sangaphandle) sesilwane esinikelayo, bese usixhuma nge-enzymatic kwenye i-molecule ye-DNA (i-cloning vector) ukuze wakhe i-molecule entsha ye-DNA.b.Dlulisela i-molecule ye-DNA ephinde ihlanganiswe kuseli lomamukeli futhi uyiphindaphinde futhi uyilondoloze kuseli lomamukeli.Le nqubo ibizwa ngokuthi ukuguqulwa.c.Khipha futhi ukhombe lawo maseli abamukeli asemunce i-DNA recombinant.d.I-massculture amaseli aqukethe i-DNA recombinant ukuthola ukuthi isakhi sosizo sangaphandle siyavezwa yini.
9. Ukwakhiwa komtapo wolwazi wezakhi zofuzo Izindlela ezintathu zokuhlolwa kwezinhlanganisela zinikeziwe futhi inqubo ichazwe kafushane.
Ukuhlolwa kokungazweli kwemithi elwa namagciwane, ukungasebenzi kokufakwa kokungazweli, ukuhlolwa kwamachashazi amhlophe okwesibhakabhaka noma ukuhlolwa kwe-PCR, ukuhlolwa okuhlukile, uphenyo lwe-DNA Iningi lama-cloning vectors lithwala izakhi zofuzo zokumelana nama-antibiotic (anti-ampicillin, tetracycline).Lapho i-plasmid idluliselwa ku-Escherichia coli, amagciwane azothola ukumelana, futhi lawo angadluliswanga ngeke akwazi ukumelana.Kodwa ayikwazi ukuhlukanisa ukuthi ihlelwe kabusha noma cha.Kuvekhtha equkethe izakhi zofuzo ezimbili zokumelana, uma isiqeshana se-DNA yangaphandle sishuthekwa kweyodwa yezakhi zofuzo futhi sibangele ukuthi isakhi sofuzo singasebenzi, izilawuli zamapuleti ezimbili eziqukethe imithi ehlukene zingasetshenziswa ukuze kuhlolwe izinhlanganisela ezinhle.Isibonelo, i-pUC plasmid iqukethe isakhi sofuzo se-LacZ (i-encoding β-galactosidase), engabolisa i-chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) ukuze ikhiqize okuluhlaza okwesibhakabhaka, ngaleyo ndlela iguqule uhlobo lube luhlaza okwesibhakabhaka.Lapho i-DNA yangaphandle ifakiwe, isakhi sofuzo se-LacZ asikwazi ukuvezwa, futhi uhlobo lumhlophe, ukuze kuhlolwe amagciwane aphinde ahlanganiswe.
10. Chaza inqubo eyisisekelo yokuthola izilwane ezishintshashintshayo ngamaseli e-embryonic stem?
Amaseli e-Embryonic stem cell (ES) angamaseli e-embryonic ngesikhathi sokuthuthukiswa kwe-embryonic, angakhuliswa ngokwenziwa futhi anwetshwe futhi abe nomsebenzi wokuhlukanisa kwezinye izinhlobo zamaseli.Isiko lamaseli e-ES: I-cell mass yangaphakathi ye-blastocyst ihlukanisiwe futhi iyakhuliswa.Uma i-ES ikhuliswa kungqimba olungaphakeli, izohlukana ibe amaseli ahlukahlukene asebenzayo njengamaseli emisipha namaseli angu-N.Uma ikhuliswe endaweni equkethe ama-fibroblasts, i-ES izogcina umsebenzi wokuhlukanisa.I-ES ingashintshwa ngofuzo, futhi umsebenzi wayo wokuhlukanisa ungahlanganiswa ngaphandle kokuthinta umsebenzi wayo wokuhlukanisa, okuxazulula inkinga yokuhlanganiswa okungahleliwe.Yethula izakhi zofuzo zangaphandle kumaseli e-embryonic stem, bese ufaka esibelethweni samagundane esifazane akhulelwe, akhule abe imidlwane, bese weqa ukuze uthole amagundane angama-homozygous.
