Bayanin ainihin sharuddan ilimin halitta
1. cDNA da cccDNA: cDNA DNA ce mai madauri biyu wanda aka haɗa ta hanyar juyar da rubutun daga mRNA;cccDNA plasmid rufaffiyar madauwari mai madauri biyu ce wacce ba ta da chromosome.
2. Standard nadawa naúrar: furotin na biyu tsarin naúrar α-helix da β-sheet iya samar da tsarin tubalan tare da na musamman geometric shirye-shirye ta daban-daban haɗa polypeptides.Irin wannan ƙayyadaddun nadawa yawanci ana kiransa tsarin super secondary.Kusan dukkan tsarin manyan makarantu ana iya siffanta su da waɗannan nau'ikan nadawa, har ma da nau'ikan nau'ikan su, don haka ana kiran su daidaitattun nadawa.
3. CAP: cyclic adenosine monophosphate (cAMP) furotin mai karɓar furotin CRP (protein mai karɓar cAMP), hadaddun da aka kafa bayan haɗuwa da cAMP da CRP ana kiran furotin mai kunnawa CAP (cAMP activated protein)
4. Jerin Palindromic: Juyin juzu'i na gaba ɗaya na wani yanki na guntun DNA, galibi wurin ƙuntatawa enzyme.
5. micRNA: Madaidaicin RNA ko antisense RNA, wanda ya dace da jerin mRNA kuma yana iya hana fassarar mRNA.
6. Ribozyme: RNA tare da aikin catalytic, wanda ke taka rawar autocatalytic a cikin tsarin splicing na RNA.
7. Motif: Akwai wasu yankuna na gida masu kama da siffa mai girma uku da topology a cikin tsarin sararin samaniya na kwayoyin furotin.
8. peptide siginar: peptide tare da ragowar amino acid 15-36 a cikin N-terminus yayin haɗin furotin, wanda ke jagorantar transmembrane na furotin.
9. Attenuator: Tsarin nucleotide tsakanin yankin mai aiki da tsarin tsarin halitta wanda ke ƙare rubutun.
10. Tabo Sihiri: Lokacin da kwayoyin cuta suka girma kuma suka gamu da rashin cikakkiyar amino acid, kwayoyin za su samar da martanin gaggawa don dakatar da bayyanar dukkan kwayoyin halitta.Alamun da ke haifar da wannan martanin gaggawa sune guanosine tetraphosphate (ppGpp) da guanosine pentaphosphate (pppGpp).Matsayin PPGpp da pppGpp ba ɗaya ba ne ko kaɗan kawai, amma yana rinjayar yawancin su, don haka ana kiran su super-regulators ko wuraren sihiri.
11. Abubuwan haɓakawa na sama: yana nufin jerin DNA wanda ke taka rawa a cikin ayyukan mai gabatarwa, kamar TATA a cikin yankin -10, TGACA a cikin yankin -35, masu haɓakawa, da masu haɓakawa.
12. Binciken DNA: wani yanki mai lakabin DNA tare da jerin sanannun, wanda aka yi amfani dashi don gano jerin abubuwan da ba a sani ba da kuma kwayar cutar da ke nunawa.
13. Jerin SD: Yana da jerin ɗaurin ribosome da mRNA, waɗanda ke tsara fassarar.
14. Monoclonal Antibody: Antibody wanda ke aiki kawai akan ƙayyadaddun antigenic guda ɗaya.
15. Cosmid: Yana da wani sinadari na DNA wanda aka gina ta wucin gadi wanda ke riƙe da yankunan COS a ƙarshen phage kuma an haɗa shi da plasmid.
16. Blue-fari tabo nunawa: LacZ gene (encoding β-galactosidase), da enzyme iya decompose chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) don samar da blue, don haka yin iri blue.Lokacin da aka shigar da DNA na waje, ba za a iya bayyana kwayar halittar LacZ ba, kuma nau'in ya zama fari, don tantance ƙwayoyin da ke sake haɗuwa.Ana kiran wannan nunin shuɗi-fari.
17. Cis-acting element: Wani takamaiman jerin tushe a cikin DNA wanda ke daidaita maganganun kwayoyin halitta.
18. Klenow enzyme: Babban guntu na DNA polymerase I, sai dai an cire aikin 5' 3' exonuclease daga DNA polymerase I holoenzyme.
19. Anchored PCR: ana amfani dashi don haɓaka DNA na sha'awa tare da jerin sanannun a ƙarshen ɗaya.An ƙara wutsiya ta poly-dG zuwa ƙarshen jerin da ba a sani ba, sannan an yi amfani da poly-dC da jerin da aka sani azaman maƙasudi don haɓaka PCR.
20. Fusion protein: Halittar furotin eukaryotic yana da alaƙa da ƙayyadaddun kwayoyin halitta, kuma sunadaran da ke tattare da fassarar asalin furotin na asali da furotin na waje yana bayyana a lokaci guda.
Sauran sharuddan ilmin halitta
1. Taswirar zahiri na DNA shine tsari wanda aka tsara guntuwar kwayoyin halittar DNA.
2. Ragewar RNase ya kasu kashi biyu (autocatalysis) da (heterocatalysis).
3. Akwai abubuwan farawa guda uku a cikin prokaryotes sune (IF-1), (IF-2) da (IF-3).
4. Sunadaran transmembrane suna buƙatar jagora (peptides sigina), kuma aikin furotin chaperones shine (taimaka don ninka sarkar peptide a cikin ƙirar asali na furotin).
5. Abubuwan da ke cikin masu talla za a iya raba su zuwa nau'i biyu: (nau'ikan masu tallatawa) da ( abubuwan gabatarwa na sama).
6. Abubuwan bincike na kwayoyin halitta sun ƙunshi sassa uku: (tsarin kwayoyin halitta), (bayanin kwayoyin halitta da tsari), da (fasaha na sake hade DNA).
7. Mahimman gwaje-gwaje guda biyu da ke nuna cewa DNA shine kwayoyin halitta sune (cututtukan pneumococcus na mice) da (T2 phage infection na Escherichia coli).m).
8. Akwai manyan bambance-bambance guda biyu tsakanin hnRNA da mRNA: (hnRNA ya rabu a cikin tsarin juyawa zuwa mRNA), (ana ƙara 5' ƙarshen mRNA tare da m7pGppp cap, kuma akwai ƙarin polyadenylation a ƙarshen 3' mRNA acid (polyA) wutsiya).
9. Abubuwan da ake amfani da su na nau'i na nau'i mai yawa na sunadaran sune (subunit hanya ce ta tattalin arziki don amfani da DNA), (zai iya rage tasirin kuskuren bazuwar a cikin haɗin furotin akan ayyukan furotin), (aikin na iya zama da kyau sosai kuma ana buɗewa da sauri kuma an rufe shi).
10. Babban abun ciki na tsarin nadawa furotin na farko ka'idar nucleation ya hada da (nucleation), (haɓaka tsarin), (sake tsarawa na ƙarshe).
11. Galactose yana da tasiri biyu akan kwayoyin cuta;a gefe guda (ana iya amfani dashi azaman tushen carbon don haɓakar tantanin halitta);a daya bangaren (shima bangaren bangon tantanin halitta ne).Sabili da haka, ana buƙatar mai haɓaka mai zaman kanta na cAMP-CRP S2 don haɗin kai na dindindin a matakin baya;a lokaci guda, ana buƙatar mai haɓaka mai dogaro da CAMP-CRP S1 don daidaita haɓakar matakin haɓaka.Rubutun yana farawa daga (S2) tare da G kuma daga (S1) ba tare da G.
12. Recombinant DNA fasahar kuma aka sani da (gene cloning) ko (molecular cloning).Babban makasudin shine (don canja wurin bayanan kwayoyin halittar DNA a cikin wata kwayar halitta zuwa wata kwayar halitta).Gwajin sake hadewar DNA na yau da kullun yana haɗa da matakai masu zuwa: (1) Cire asalin halittar da aka yi niyya (ko ƙwayar cuta ta waje) na kwayar halitta mai ba da gudummawa, kuma a haɗa ta ta hanyar enzymatic zuwa wani kwayar halittar DNA (cloning vector) don samar da sabon kwayar halittar DNA mai sake haɗawa.② Ana canza kwayar halittar DNA ta sake haɗawa zuwa cikin tantanin halitta mai karɓa kuma ana yin shi a cikin tantanin halitta mai karɓa.Ana kiran wannan tsari canji.③ Allon allo kuma gano waɗancan sel masu karɓa waɗanda suka sha recombinant DNA.④ Haɓaka ƙwayoyin da ke ɗauke da DNA recombinant a cikin adadi mai yawa don gano ko an bayyana kwayoyin taimakon agaji na ƙasashen waje.
13. Akwai nau'ikan nau'ikan plasmid iri biyu: waɗanda ke da ƙarfi sosai ta hanyar haɗin furotin cell ana kiran su (m plasmids), kuma waɗanda ba a sarrafa su ta hanyar haɗin furotin cell ɗin ana kiran su (lalata plasmids).
14. Tsarin amsawar PCR yakamata ya kasance yana da waɗannan sharuɗɗan: a.Matsalolin DNA (kimanin tushe guda 20) tare da madaidaitan jeri a kowane ƙarshen igiyoyi biyu na kwayar halittar da za a raba.b.Enzymes tare da kwanciyar hankali na thermal kamar: TagDNA polymerase.c, dNTPd, jerin DNA na sha'awa azaman samfuri
15. Tsarin amsawa na asali na PCR ya haɗa da matakai uku: (denaturation), (annealing), da (tsawo).
16. Tsarin asali na dabbobin transgenic yawanci ya haɗa da: ① Gabatar da kwayar halitta ta waje ta cloned a cikin tsakiya na kwai da aka haɗe ko embryonic stem cell;②Dasawar kwai da aka yi wa hadi ko tantanin tantanin halitta zuwa mahaifar mace;③Cikakken ci gaban amfrayo da girma Ga 'ya'yan da ke da kwayoyin halitta na waje;④ Yi amfani da waɗannan dabbobin da za su iya samar da furotin na waje a matsayin kayan kiwo don haifar da sababbin layin homozygous.
17. Hybridoma cell Lines an samar da su ta hanyar hybridizing (spleen B) kwayoyin halitta tare da (myeloma), kuma tun da (kwayoyin ƙwayoyin cuta) zasu iya amfani da hypoxanthine da (kwayoyin kasusuwa) suna samar da ayyukan rarraba kwayoyin halitta, ana iya girma a cikin HAT matsakaici.girma.
18. Tare da zurfafa bincike, ana kiran ƙarni na farko na rigakafi (antibodies polyclonal), ƙarni na biyu (antibodies monoclonal), da ƙarni na uku (maganin injiniyan ƙwayoyin cuta).
19. A halin yanzu, aikin injiniya na ƙwayoyin cuta na ƙwayoyin cuta ya fi mayar da hankali ga baculovirus, wanda aka bayyana a cikin gabatarwar (exogenous toxin gene);(kwayoyin halittar da ke rushe tsarin rayuwar kwari na yau da kullun);(gyaran kwayoyin halittar ƙwayoyin cuta).
20. Abubuwan furotin da ke aiki da su daidai da abubuwan gama gari TATA, GC, da CAAT a cikin mammalian RNA polymerase II mai gabatarwa sune (TFIID), (SP-1) da (CTF/NF1), bi da bi.
ashirin da daya.Abubuwan da aka rubuta na asali na RNA polymerase Ⅱ sune, TFⅡ-A, TFⅡ-B, TFII-D, TFⅡ-E, kuma jerin daurin su shine: (D, A, B, E).Inda aikin TFII-D yake (daure zuwa akwatin TATA).
ashirin da biyu.Yawancin abubuwan rubutun da ke ɗaure ga DNA suna aiki ta hanyar dimers.Wuraren aiki na abubuwan rubutawa waɗanda ke ɗaure ga DNA galibi sune masu biyowa (helix-turn-helix), (motif ɗin yatsa zinc), (basic-leucine) motif zipper).
ashirin da uku.Akwai nau'ikan ƙuntatawa guda uku: (Yanke a cikin 'gefen Symmetry na sama don samar da madaidaicin madaidaici don samar da madaidaitan sassan).
ashirin da hudu.Plasmid DNA yana da sigogi daban-daban guda uku: (tsarin SC), (oc configuration), (tsarin L).Na farko a cikin electrophoresis shine (SC configuration).
25. Exogenous gene expression tsarin, yafi (Escherichia coli), (Yast), (Insect) da (mammalian cell tebur).
26. Hanyoyin da aka saba amfani da su don dabbobin transgenic sune: (hanyar kamuwa da cuta ta retroviral), (Hanya microinjection DNA), (hanyar embryonic stem cell).
Aikace-aikacen Halittar Halitta
1. Sunan ayyukan RNA fiye da 5?
Canja wurin RNA tRNA Amino acid Ribosome RNA rRNA Ribosome ya ƙunshi manzo RNA mRNA Protein kira samfuri Samfuran Nukiliya mai ban sha'awa RNA hnRNA Mafarin balagagge mRNA ƙaramin nukiliyar RNA snRNA Ya shiga cikin hnRNA splicing Ƙananan cytoplasmic RNA scRNA/7SL-RNA furotin RNA SRNA / 7SL-RNA furotin da aka yi da siginar siginar RNA da aka yi da siginar DNA da aka yi da shi Ribozyme RNA Enzymatically aiki RNA
2. Menene babban bambanci tsakanin masu tallata prokaryotic da eukaryotic?
Prokaryotic TTGACA --- TATAAT------Shafin Ƙaddamarwa-35 -10 Eukaryotic Enhancer---GC ---CAAT---TAATAA-5mGpp-Shafin Ƙaddamarwa-110 -70 -25
3. Menene babban al'amurran gina wucin gadi na plasmids na halitta?
Plasmids na halitta sau da yawa suna da lahani, don haka ba su dace da amfani da su azaman masu ɗaukar nauyin injiniyan kwayoyin halitta ba, kuma dole ne a gyara su kuma a gina su: a.Ƙara kwayoyin halitta masu alamar zaɓi masu dacewa, kamar biyu ko fiye, waɗanda suke da sauƙin amfani don zaɓi, yawanci kwayoyin ƙwayoyin cuta.b.Ƙara ko rage wuraren yankan enzyme masu dacewa don sauƙaƙe haɗuwa.c.Rage tsayin daka, yanke ɓangarorin da ba dole ba, inganta haɓakar shigo da kayayyaki da haɓaka ƙarfin lodi.d.Canja kwafi, daga matsi zuwa sako-sako, daga ƴan kwafi zuwa ƙarin kwafi.e.Ƙara abubuwa na musamman na kwayoyin halitta bisa ga buƙatun musamman na injiniyan kwayoyin halitta
4. Ba da misali na hanya don tantance bambancin cDNA na musamman na nama?
An shirya yawan tantanin halitta guda biyu, ana bayyana kwayar halittar da aka yi niyya ko kuma an bayyana su sosai a cikin ɗayan sel, kuma ba a bayyana kwayar da aka yi niyya ko ƙasƙantar da ita a cikin ɗayan tantanin halitta ba, sannan ana samun nau'in manufa ta hanyar haɓakawa da kwatantawa.Misali, yayin faruwa da ci gaban ciwace-ciwacen ciwace-ciwacen ciwace-ciwacen ciwace-ciwace, kwayoyin tumor za su gabatar da mRNAs tare da matakan magana daban-daban fiye da sel na yau da kullun.Don haka, ana iya tantance kwayoyin halittar da ke da alaƙa da ƙari ta hanyar haɓakawa daban-daban.Hakanan za'a iya amfani da hanyar ƙaddamarwa don tantance kwayoyin halittar da aka jawo bayyanar su.
5. Ƙirƙiri da nunawa na layin salula na hybridoma?
Kwayoyin ƙwayoyin cuta na B + ƙwayoyin myeloma, ƙara polyethylene glycol (PEG) don haɓaka haɗin sel, da ƙwayoyin ƙwayoyin cuta na B-myeloma da ke girma a cikin matsakaicin HAT (wanda ke ɗauke da hypoxanthine, aminopterin, T) suna ci gaba da haɓaka abinci mai gina jiki.Fusishin tantanin halitta ya ƙunshi: sel mai ban tsoro mai ban tsoro: ba za a iya yin girma ba, ba za a iya warware su ba, ba za a iya warware su ba, ba za a iya warware su a cikin Vitro ba.Kwayoyin haɗakar kashi-kashi: ba za su iya amfani da hypoxanthine ba, amma suna iya haɗa purine ta hanya ta biyu ta amfani da folate reductase.Aminopterin yana hana folate reductase don haka ba zai iya girma ba.Kwayoyin fusion na kasusuwa: na iya girma a cikin HAT, ƙwayoyin splin na iya amfani da hypoxanthine, kuma ƙwayoyin kasusuwa suna ba da aikin rarraba tantanin halitta.
6. Menene ka'ida da hanyar ƙayyade ainihin tsarin DNA ta hanyar dideoxy ta ƙarshe (hanyar Sanger)?
Ka'idar ita ce a yi amfani da mai ƙare sarkar nucleotide-2,,3,-dideoxynucleotide don ƙare tsawo na DNA.Tun da ba shi da 3-OH da ake buƙata don samuwar 3/5/phosphodiester bond, da zarar an haɗa shi cikin sarkar DNA, ba za a iya ƙara sarkar DNA ba.Bisa ga ka'idar haɗin ginin tushe, a duk lokacin da DNA polymerase ke buƙatar dNMP don shiga cikin sarkar DNA ta yau da kullum, akwai hanyoyi guda biyu, daya shine shiga cikin ddNTP, wanda ke haifar da ƙarewar deoxynucleotide sarkar tsawo;ɗayan shine shiga cikin dNTP , ta yadda sarkar DNA zata iya ci gaba da tsawo har sai an haɗa ddNTP na gaba.Bisa ga wannan hanyar, ana iya samun rukuni na guntun DNA na tsayi daban-daban da ke ƙarewa a ddNTP.Hanyar ita ce a rarraba zuwa ƙungiyoyi huɗu bi da bi ddAMP, ddGMP, ddCMP, da ddTMP.Bayan amsawa, polyacrylamide gel electrophoresis na iya karanta jerin DNA bisa ga makada na iyo.
7. Menene ingantacciyar tasirin ƙa'ida ta furotin mai kunnawa (CAP) akan rubutun?
Cyclic adenylate (cAMP) furotin mai karɓa na CRP (protein mai karɓar cAMP), hadaddun da aka kafa ta hanyar haɗin cAMP da CRP ana kiransa CAP (cAMPactivated protein).Lokacin da E. coli ke girma a cikin matsakaicin rashin glucose, haɗin CAP yana ƙaruwa, kuma CAP yana da aikin kunna masu tallata kamar lactose (Lac).Wasu masu tallata masu dogaro da CRP ba su da fasalin fasalin yanki -35 na yau da kullun (TTGACA) waɗanda masu tallata gama gari suke da shi.Don haka, yana da wahala RNA polymerase ta ɗaure ta.Kasancewar CAP (aiki): na iya inganta haɓakar daurin enzyme da mai haɓakawa sosai.Yafi nuna abubuwa guda biyu masu zuwa: ① CAP yana taimakawa kwayoyin enzyme don daidaitawa daidai ta hanyar canza yanayin mai gabatarwa da hulɗa tare da enzyme, don haɗuwa tare da yankin -10 kuma yana taka rawar maye gurbin aikin yankin -35.②CAP kuma na iya hana daurin RNA polymerase zuwa wasu shafuka a cikin DNA, ta haka yana ƙara yuwuwar ɗaure ga takamaiman mai tallata sa.
8. Waɗanne matakai yawanci ake haɗawa a cikin gwajin sake haɗa DNA na yau da kullun?
a.Ciro asalin halittar da aka yi niyya (ko exogenous gene) na kwayar halitta mai ba da gudummawa, kuma a haɗa ta da enzymatically zuwa wani kwayar halittar DNA (cloning vector) don samar da sabon kwayar halittar DNA ta sake haɗawa.b.Canja wurin kwayar halittar DNA mai sake haɗewa zuwa cikin tantanin halitta mai karɓa kuma a yi kwafi kuma adana shi a cikin tantanin halitta mai karɓa.Ana kiran wannan tsari canji.c.Allon allo kuma gano waɗancan sel masu karɓa waɗanda suka mamaye DNA ɗin da ke sake haɗawa.d.Al'adar taro sel ɗin da ke ɗauke da DNA ɗin sake haɗewa don gano ko an bayyana kwayoyin taimakon agaji na ƙasashen waje.
9. Gina laburaren kwayoyin halitta an ba da hanyoyi uku don tantance abubuwan sake haɗawa kuma an bayyana tsarin a taƙaice.
Gwajin juriya na ƙwayoyin cuta, rashin kunna juriya na sakawa, nunin tabo mai launin shuɗi ko duban PCR, tantancewa daban-daban, binciken DNA Yawancin ƙwayoyin cuta na ƙwayoyin cuta suna ɗauke da kwayoyin juriya na ƙwayoyin cuta (anti-ampicillin, tetracycline).Lokacin da plasmid aka canjawa wuri zuwa Escherichia coli, kwayoyin za su sami juriya, kuma wadanda ba tare da canja wuri ba za su sami juriya.Amma ba zai iya tantance ko an sake tsara shi ko a'a.A cikin na'urar da ke dauke da kwayoyin halitta na juriya guda biyu, idan aka saka wani guntun DNA na waje a cikin daya daga cikin kwayoyin halitta kuma ya sa kwayar ta daina aiki, ana iya amfani da sarrafa faranti guda biyu da ke dauke da magunguna daban-daban don tantance abubuwan da suka dace.Alal misali, pUC plasmid ya ƙunshi nau'in LacZ (encoding β-galactosidase), wanda zai iya lalata chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) don samar da shuɗi, don haka juya launin shuɗi.Lokacin da aka shigar da DNA na kasashen waje, ba za a iya bayyana kwayar halittar LacZ ba, kuma nau'in ya zama fari, don tantance kwayoyin da ke sake hadewa.
10. Bayyana ainihin tsari na samun dabbobi masu canzawa ta hanyar kwayoyin halitta na amfrayo?
Kwayoyin da ke cikin mahaifa (ES) su ne ƙwayoyin amfrayo yayin haɓakar amfrayo, waɗanda za a iya haɓaka ta hanyar wucin gadi da haɓaka kuma suna da aikin bambanta zuwa wasu nau'in sel.Al'adar Kwayoyin ES: Yawan tantanin halitta na ciki na blastocyst ya keɓanta da al'ada.Lokacin da aka yi al'adar ES a cikin wani nau'in mai ba da abinci, zai bambanta zuwa sel masu aiki daban-daban kamar ƙwayoyin tsoka da ƙwayoyin N.Lokacin da aka haɓaka a cikin matsakaici mai ɗauke da fibroblasts, ES zai kula da aikin bambanta.Ana iya sarrafa ES ta hanyar kwayoyin halitta, kuma ana iya haɗa aikin bambancinsa ba tare da shafar aikin bambancinsa ba, wanda ke magance matsalar haɗin kai bazuwar.Gabatar da kwayoyin halittar da ba a iya gani ba a cikin kwayoyin halitta na amfrayo, sannan a dasa su a cikin mahaifar berayen mata masu juna biyu, su zama ’ya’ya, su ketare don samun berayen homozygous.
