Ibisobanuro byamagambo yibanze ya biologiya
1. cDNA na cccDNA: cDNA ni ADN igizwe n'imirongo ibiri ikomatanyirijwe hamwe na transcriptase iva muri mRNA;cccDNA ni plasmid ya kabiri-ifunze uruziga ruzengurutse ADN idafite chromosome.
2. Igikoresho gisanzwe: poroteyine yubaka icyiciro cya kabiri α-helix na β-urupapuro rushobora gukora ibice byubaka hamwe na geometrike idasanzwe binyuze muri polypeptide zitandukanye.Ubu bwoko bwagenwe bukunze kwitwa super secondary structure.Inzira hafi ya zose zo murwego zishobora gusobanurwa nubwoko bwikubye, ndetse nubwoko bwazo, bityo rero byitwa ibice bisanzwe byububiko.
3. CA
4. Urukurikirane rwa Palindromic: Urukurikirane rwuzuzanya rwurwego rwigice cya ADN, akenshi urubuga rwa enzyme rubuza.
5. micRNA: Kwivanga kwuzuye RNA cyangwa antisense RNA, byuzuzanya nurutonde rwa mRNA kandi bishobora kubuza guhindura mRNA.
6. Ribozyme: RNA hamwe nibikorwa bya catalitiki, bigira uruhare rwa autocatalytic mugikorwa cyo gutera RNA.
7. Motif: Hariho uturere tumwe na tumwe dufite imiterere-yuburyo butatu na topologiya muburyo butandukanye bwa molekile ya poroteyine
8. Peptide yerekana ibimenyetso: peptide ifite ibisigisigi bya aside amine 15-36 kuri N-terminus mugihe cya synthesis ya protein, iyobora transembrane ya proteine.
9. Attenuator: Urutonde rwa nucleotide hagati yakarere gakorera hamwe na gene yimiterere ihagarika transcript.
10. Umwanya wa Magic: Iyo bagiteri ikuze igahura no kubura burundu aside amine, bagiteri izatanga igisubizo cyihutirwa kugirango ihagarike imvugo ya genes zose.Ibimenyetso bitanga igisubizo cyihutirwa ni guanosine tetraphosphate (ppGpp) na pentaphosphate ya guanosine (pppGpp).Uruhare rwa PpGpp na pppGpp ntabwo ari umwe cyangwa bake bakora gusa, ahubwo bigira ingaruka kumubare munini wabyo, bityo bakitwa super-regulator cyangwa ahantu h'ubumaji.
11. Upstream promoter element: bivuga urutonde rwa ADN rufite uruhare runini mugikorwa cya promoteri, nka TATA mukarere ka -10, TGACA mukarere ka -35, abazamura, hamwe na attenuator.
12. Ubushakashatsi bwa ADN: igice cyanditseho ADN hamwe nurutonde ruzwi, rukoreshwa cyane mugutahura urutonde rutazwi hamwe na gen zigamije.
13. Urutonde rwa SD: Nuburyo bukurikirana bwa ribosome na mRNA, bigenga ubusobanuro.
14. Antibody ya Monoclonal: Antibody ikora kurwanya antigenic imwe gusa.
15. Cosmid: Nuburyo bwububiko bwububiko bwububiko bwa ADN bugumana uturere twa COS kumpande zombi zicyiciro kandi buhujwe na plasmid.
16. Kugaragaza ibibara byera-byera: LacZ gene (encoding β-galactosidase), enzyme irashobora kubora chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kugirango itange ubururu, bityo itume ubururu buba ubururu.Iyo ADN idasanzwe yinjizwemo, gene ya LacZ ntishobora kugaragazwa, kandi umutwaro wera, kugirango ugaragaze bagiteri za recombinant.Ibi byitwa ubururu-bwera bwerekana.
17. Ikintu gikora ibintu: Urutonde rwihariye rwibanze muri ADN igenga imiterere ya gene.
18. Enzyme ya Klenow: Igice kinini cya ADN polymerase ya I, usibye ko ibikorwa bya 5 '3' exonuclease byakuwe muri ADN polymerase I holoenzyme
19. Anchored PCR: ikoreshwa mugukomeza ADN yinyungu hamwe nurutonde ruzwi kuruhande rumwe.Umurizo wa poly-dG wongeyeho kumpera imwe yuruhererekane rutazwi, hanyuma poly-dC hamwe nurutonde ruzwi byakoreshejwe nka primers yo kongera PCR.
20. Poroteyine ya Fusion: Gene ya poroteyine ya eukaryotique ifitanye isano na gene ya exogenous, kandi poroteyine igizwe no guhindura poroteyine y’umwimerere na poroteyine ya exogenous igaragarira icyarimwe.
Andi magambo ya biologiya
1. Ikarita ifatika ya ADN ni gahunda itondekanya (kubuza endonuclease-digested) ibice bya molekile ya ADN.
2. Igice cya RNase kigabanyijemo ubwoko bubiri (autocatalyse) na (heterocatalyse).
3. Hariho ibintu bitatu byo gutangiza muri prokaryote ni (IF-1), (IF-2) na (IF-3).
4. Poroteyine za Transmembrane zisaba ubuyobozi (ibimenyetso peptide yerekana), kandi uruhare rwa protein chaperone ni (ifasha kuzinga urunigi rwa peptide muburyo bwa poroteyine).
5. Ibintu muri porotokoro birashobora kugabanywa muburyo bubiri: (ibyingenzi byamamaza) hamwe na (upstream promoter element).
6. Ibirimo mubushakashatsi bwibinyabuzima bya molekuline bikubiyemo ahanini ibice bitatu: (ibinyabuzima byubaka imiterere), (imvugo ya gene nubuyobozi), na (tekinoroji ya ADN).
7. Ubushakashatsi bubiri bwingenzi bwerekana ko ADN ari genetike ni (kwandura pneumococcus yimbeba) na (T2 fage infection ya Escherichia coli).ubushobozi).
8. Hariho itandukaniro ryibanze bibiri hagati ya hnRNA na mRNA: (hnRNA itangwa mugikorwa cyo guhinduka muri mRNA), (5 'iherezo rya mRNA ryongewemo numutwe wa m7pGppp, kandi hariho polyadenylation yinyongera kuri 3' iherezo ryumurizo wa mRNA (polyA)).
9. Ibyiza byuburyo bwinshi bwa subunit ya proteine ni (subunit nuburyo bwubukungu bwo gukoresha ADN), (irashobora kugabanya ingaruka zamakosa yatanzwe muri synthesis ya protein kubikorwa bya poroteyine), (ibikorwa birashobora gufungurwa neza kandi byihuse bikingurwa kandi bigafungwa).
10. Ibyingenzi byingenzi byububiko bwa poroteyine uburyo bwa mbere bwa nucleation harimo (nucleation), (gutunganyiriza imiterere), (rearrangement ya nyuma).
11. Galactose igira ingaruka ebyiri kuri bagiteri;kuruhande rumwe (irashobora gukoreshwa nkisoko ya karubone yo gukura kwingirabuzimafatizo);kurundi ruhande (ni nacyo kigize urukuta rw'akagari).Kubwibyo, cAMP-CRP-yigenga iteza imbere S2 irakenewe kugirango synthesis ihoraho kurwego rwinyuma;icyarimwe, cAMP-CRP-iterwa na porotokoro S1 irakenewe kugirango igenzure urwego rwohejuru.Kwandukura bitangirira kuri (S2) hamwe na G no kuva (S1) nta G.
12. Tekinoroji ya ADN ya recombinant izwi kandi nka (cloni ya gene) cyangwa (clone ya molekile).Intego nyamukuru ni (guhererekanya amakuru yimiterere ADN mubinyabuzima bumwe mubindi binyabuzima).Ubushakashatsi busanzwe bwa ADN bwo kwisubiramo busanzwe bukubiyemo intambwe zikurikira: (1) Kuramo gene igenewe (cyangwa gene exogenous) yibinyabuzima byabaterankunga, hanyuma uyihuze na enterineti na molekile ya ADN (clonctor vector) kugirango ikore molekile nshya ya ADN ya recombinant.Molecu molekile ya ADN ya recombinant yimurirwa mu kagari yakiriye kandi ikopororwa mu kagari kahawe.Iyi nzira yitwa guhinduka.③ Erekana kandi umenye utugingo ngengabuzima twakiriye ADN ya recombinant.UltKora selile zirimo ADN ya recombinant ku bwinshi kugirango umenye niba gene y’amahanga itangwa.
13. Hariho ubwoko bubiri bwo kwigana plasmide: izigenzurwa cyane na synthesis ya proteine ya selile yakira yitwa (plasmide ifatanye), naho ibitagenzurwa cyane na synthesis ya proteine selile byitwa (plasmide yoroheje).
14. Sisitemu ya reaction ya PCR igomba kuba ifite ibi bikurikira: a.ADN yibanze (hafi 20 shingiro) hamwe nuburyo bwuzuzanya kuri buri mpera yimirongo ibiri ya gen igenewe gutandukana.b.Enzymes hamwe nubushyuhe bwumuriro nka: TagDNA polymerase.c, dNTPd, ADN ikurikirana yinyungu nkicyitegererezo
15. Igikorwa cyibanze cyibikorwa bya PCR gikubiyemo ibyiciro bitatu: (denaturation), (annealing), na (kwagura).
16. Inzira y'ibanze y’inyamaswa zanduye zikubiyemo:Gutera amagi yatewe intanga cyangwa ingirangingo ngengabuzima mu nda ibyara;Development Iterambere ryuzuye ryo gusama no gukura Kubyara hamwe na genes zamahanga;These Koresha inyamaswa zishobora kubyara poroteyine zo mu mahanga nk'ubworozi bwo korora imirongo mishya itaryamana.
17. Imirongo ya selile ya Hybridoma ikorwa no kuvanga (spleen B) selile hamwe na (myeloma), kandi kubera ko (selile spleen selile) ishobora gukoresha hypoxanthine kandi (selile amagufa) itanga imirimo yo kugabana selile, irashobora gukura muburyo bwa HAT.gukura.
18. Hamwe nubushakashatsi bwimbitse, igisekuru cya mbere cya antibodi cyitwa (antibodiyite ya polyclone), igisekuru cya kabiri (antibodiyite za monoclonal), nigisekuru cya gatatu (antibodies genetique).
19. Kugeza ubu, ingengabihe ya virusi y’udukoko yibanda cyane cyane kuri baculovirus, igaragarira mu kwinjiza (gene exogenous toxin);(ingirabuzimafatizo zibangamira ubuzima busanzwe bw'udukoko);(guhindura genes za virusi).
20. Impinduka za poroteyine zikora zijyanye nibintu bisanzwe TATA, GC, na CAAT mumatungo yinyamabere RNA polymerase II yamamaza ni (TFIID), (SP-1) na (CTF / NF1).
makumyabiri na rimwe.Impamvu zingenzi zo kwandukura za RNA polymerase Ⅱ ni, TFⅡ-A, TFⅡ-B, TFII-D, TFⅡ-E, kandi uko bikurikirana ni: (D, A, B, E).Aho imikorere ya TFII-D ari (guhuza agasanduku ka TATA).
makumyabiri na kabiri.Byinshi mubintu byandikirwa bihuza ADN ikora muburyo bwa dimers.Imikorere yimikorere yibintu byandikirwa bihuza ADN mubisanzwe ni ibi bikurikira (helix-turn-helix), (zinc urutoki motif), (shingiro-leucine) zipper motif).
makumyabiri na gatatu.Hariho ubwoko butatu bwo kubuza endonuclease clavage uburyo: (gukata kuruhande rwa 5 'kuruhande rwa simmetrie axis kugirango ubyare 5' bifatanye), (gabanya kuruhande rwa 3 'kuruhande rwa simmetry axis kugirango ubyare 3' bifatanye (gukata kumurongo wa simmetrie kugirango ubyare ibice).
makumyabiri na bane.ADN ya Plasmid ifite ibice bitatu bitandukanye: (Iboneza rya SC), (iboneza rya oc), (L iboneza).Iyambere muri electrophoreis ni (iboneza rya SC).
25. Sisitemu yo kwerekana imiterere ya gene idasanzwe, cyane cyane (Escherichia coli), (Umusemburo), (Udukoko) na (Imbonerahamwe y'inyamabere).
26. Uburyo bukunze gukoreshwa ku nyamaswa zanduye ni: (uburyo bwo kwandura virusi itera virusi), (uburyo bwa microinjection ya ADN), (uburyo bwa embryonic stem selile).
Gushyira mu bikorwa ibinyabuzima
1. Vuga imikorere ya RNA zirenga 5?
Kwimura RNA tRNA Kwimura aside amine Ribosome RNA rRNA Ribosome igizwe nintumwa RNA mRNA Protein synthesis template Heterogeneous nuclear RNA hnRNA Precursor ya mRNA ntoya ya kirimbuzi RNA snRNA Ifite uruhare muri hnRNA itera cytoplazimike RNA scRNA / 7SL-RNA protein Plasma reticulum-yerekana neza umubiri wa RNA.
2. Ni irihe tandukaniro nyamukuru riri hagati ya porokaryyotike na eukaryotic?
Prokaryotic TTGACA --- TATAAT ------ Urubuga rwo gutangiza-35 -10 Umuyoboro wa Eukaryotic --- GC --- CAAT ---- TATAA-5mGpp-Urubuga rwo gutangiza-110 -70 -25
3. Ni ubuhe buryo bw'ingenzi bwubaka ibihimbano bya plasima karemano?
Plasima karemano ikunze kugira inenge, ntabwo rero ikwiriye gukoreshwa nkabatwara ubwubatsi bwa geneti, kandi igomba guhinduka ikubakwa: a.Ongeramo ibimenyetso byerekana ibimenyetso bifatika, nka bibiri cyangwa byinshi, byoroshye gukoresha muguhitamo, mubisanzwe antibiotique.b.Ongera cyangwa ugabanye urubuga rukwiye rwa enzyme kugirango byorohereze.c.Gabanya uburebure, gabanya ibice bitari ngombwa, kunoza imikorere yo gutumiza no kongera ubushobozi bwo gupakira.d.Hindura kopi, kuva kumurongo kugeza kurekuye, kuva kopi nkeya kugeza kuri kopi nyinshi.e.Ongeramo ibintu byihariye bya genetike ukurikije ibisabwa byihariye bya injeniyeri
4. Tanga urugero rwuburyo bwo kwerekana itandukaniro rya cDNA yihariye?
Abantu babiri b'utugari barateguwe, gene igenewe iragaragazwa cyangwa igaragazwa cyane muri imwe mu ngirabuzimafatizo, kandi gene igenewe ntigaragazwa cyangwa ngo igaragare mu rundi rugari, hanyuma gene igenewe ibonwa no kuvanga no kugereranya.Kurugero, mugihe cyo kubaho no gukura kw'ibibyimba, selile yibibyimba izerekana mRNAs hamwe nurwego rwimvugo zitandukanye kuruta selile zisanzwe.Kubwibyo, genes zijyanye nibibyimba zirashobora kugenzurwa no kuvanga ibintu bitandukanye.Uburyo bwa induction burashobora kandi gukoreshwa mugusuzuma gen zigaragaza imvugo.
5. Igisekuru no kwerekana imirongo ya selile ya hybridoma?
Spleen B selile + myeloma selile, ongeramo polyethylene glycol (PEG) kugirango uteze imbere ingirabuzimafatizo, hamwe na selile B-myeloma fusion selile ikura muburyo bwa HAT (irimo hypoxanthine, aminopterine, T) ikomeza kwagura intungamubiri.Guhuza ingirabuzimafatizo birimo: spleen-spleen fusion selile: idashobora gukura, ingirabuzimafatizo ntishobora guterwa muri vitro.Amagufa-amagufwa yo guhuza ingirabuzimafatizo: ntashobora gukoresha hypoxanthine, ariko arashobora guhuza purine binyuze munzira ya kabiri ukoresheje folate reductase.Aminopterine ibuza folate reductase bityo ntishobora gukura.Amagufa-spleen fusion selile: arashobora gukura muri HAT, selile spleen selile irashobora gukoresha hypoxanthine, naho selile igufa itanga imikorere yo kugabana selile.
6. Ni irihe hame n'uburyo bwo kumenya imiterere y'ibanze ya ADN hakoreshejwe uburyo bwo kurangiza dideoxy (uburyo bwa Sanger)?
Ihame ni ugukoresha nucleotide yumunyururu-2 ,, 3, -dideoxynucleotide kugirango uhagarike kwaguka kwa ADN.Kubera ko ibuze 3-OH isabwa kugirango habeho imigozi ya 3/5 / fosifori, iyo imaze kwinjizwa mumurongo wa ADN, urunigi rwa ADN ntirushobora kwagurwa.Ukurikije ihame ryo guhuza shingiro, igihe cyose ADN polymerase ikeneye dNMP kugirango igire uruhare muburyo busanzwe bwagutse bwa ADN, haribintu bibiri bishoboka, kimwe nukwitabira ddNTP, bikaviramo guhagarika kwagura urunigi rwa deoxynucleotide;ikindi nukwitabira dNTP, kugirango urunigi rwa ADN rushobora gukomeza kwaguka kugeza ddNTP itaha.Ukurikije ubu buryo, itsinda rya ADN ibice byuburebure butandukanye burangirira kuri ddNTP birashobora kuboneka.Uburyo nugucamo amatsinda ane ddAMP, ddGMP, ddCMP, na ddTMP.Nyuma yo kubyitwaramo, polyacrylamide gel electrophoreis irashobora gusoma urutonde rwa ADN ukurikije imirongo yo koga.
7. Ni izihe ngaruka nziza zo kugenzura poroteyine ikora (CAP) kuri transcript?
Cyclic adenylate (cAMP) reseptor protein CRP (cAMP reseptor protein), uruganda rwakozwe no guhuza cAMP na CRP rwitwa CAP (proteine cAMPactivated).Iyo E. coli ikuze muburyo butagira glucose, synthesis ya CAP iriyongera, kandi CAP ifite umurimo wo gukora porotokoro nka lactose (Lac).Bamwe mubateza imbere CRP babuze uburyo busanzwe -35 bwakurikiranye mukarere (TTGACA) abamamaza basanzwe bafite.Kubwibyo, biragoye kuri RNA polymerase kuyihuza nayo.Kubaho kwa CAP (imikorere): birashobora kunoza cyane guhuza guhuza enzyme na poroteri.Yerekana cyane cyane ibintu bibiri bikurikira: AP CAP ifasha molekile ya enzyme kwerekera neza muguhindura ihinduka rya porotokoro ndetse n’imikoranire na enzyme, kugirango uhuze nakarere ka -10 kandi ugire uruhare rwo gusimbuza imikorere yakarere ka -35.APCAP irashobora kandi kubuza guhuza polymerase ya RNA ku zindi mbuga muri ADN, bityo bikongerera amahirwe yo guhuza na porotokoro yihariye.
8. Ni izihe ntambwe zikunze gushyirwa mubushakashatsi bwa ADN busanzwe?
a.Kuramo gene igenewe (cyangwa exogenous gene) yibinyabuzima byabaterankunga, hanyuma ubihuze muyindi mikorere ya molekile ya ADN (clonctor vector) kugirango ukore molekile nshya ya ADN ya recombinant.b.Hindura molekile ya ADN ya recombinant muri selile yakiriye hanyuma uyigane kandi uyibike muri selile yakiriye.Iyi nzira yitwa guhinduka.c.Erekana kandi umenye utugingo ngengabuzima twakiriye ADN ya recombinant.d.Umuco rusange ingirabuzimafatizo zirimo ADN ya recombinant kugirango tumenye niba gene yimfashanyo zamahanga zagaragajwe.
9. Kubaka isomero rya gene Uburyo butatu bwo gusuzuma recombinants butangwa kandi inzira irasobanuwe muri make.
Kugenzura antibiyotike yo kurwanya, kudashyiramo insimburangingo yo kurwanya, kwerekana ibara ry'ubururu-bwera cyangwa kwerekana PCR, gusuzuma itandukaniro, ubushakashatsi bwa ADN Ubushakashatsi bwinshi bwa cloni butwara genes zo kurwanya antibiyotike (anti-ampicillin, tetracycline).Iyo plasmid yimuriwe muri Escherichia coli, bagiteri izabona imbaraga, kandi abadafite transfert ntibazagira imbaraga.Ariko ntishobora gutandukanya niba yaravuguruwe cyangwa idahinduwe.Muri vector irimo genes ebyiri zo kurwanya, niba igice cya ADN cyo mumahanga cyinjijwe muri imwe muri gen kandi bigatuma gene idakora, igenzura rya plaque ebyiri zirimo imiti itandukanye irashobora gukoreshwa mugupima recombinants nziza.Kurugero, plasmid ya pUC irimo gene ya LacZ (encoding β-galactosidase), ishobora kubora chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kugirango itange ubururu, bityo ihindure ubururu.Iyo ADN y’amahanga yinjizwemo, gene ya LacZ ntishobora kugaragazwa, kandi umutwaro wera, kugirango ugaragaze bagiteri za recombinant.
10. Sobanura inzira y'ibanze yo kubona inyamaswa zanduye binyuze mu ngirabuzimafatizo?
Embryonic stem selile (ES) ni ingirangingo zintangangore mugihe cyo gukura kwa emboro, zishobora kuba imico yubukorikori kandi ikagwira kandi ikagira umurimo wo gutandukanya ubundi bwoko bwa selile.Umuco w'utugingo ngengabuzima twa ES: Imbere ya selile y'imbere ya blastocyst iri mu bwigunge kandi ifite umuco.Iyo ES itezimbere murwego rutagira ibiryo, bizatandukana mubice bitandukanye bikora nka selile yimitsi na selile N.Iyo itezimbere muburyo burimo fibroblast, ES izakomeza imikorere itandukanye.ES irashobora gukoreshwa muburyo bwa genetike, kandi imikorere yayo yo gutandukanya irashobora guhuzwa bitagize ingaruka kumikorere yayo itandukanye, ikemura ikibazo cyo kwishyira hamwe.Injira ingirabuzima fatizo mu ngirabuzimafatizo ngengabuzima, hanyuma ushyire muri nyababyeyi y’imbeba z’abagore batwite, ukure mu bibwana, hanyuma wambuke kugirango ubone imbeba zidahuje igitsina.