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Kufotokozera mawu a Basic Molecular Biology

Zida za Molecular Biology

1. cDNA ndi cccDNA: cDNA ndi DNA yamitundu iwiri yopangidwa ndi reverse transcriptase kuchokera ku mRNA;cccDNA ndi DNA yozungulira yozungulira yozungulira iwiri ya plasmid yopanda chromosome.
2. Chigawo chopinda chokhazikika: puloteni yachiwiri ya kapangidwe ka α-helix ndi β-sheet ikhoza kupanga midadada yopangidwa ndi makonzedwe apadera a geometric kupyolera mu ma polypeptides osiyanasiyana ogwirizanitsa.Kupinda kotsimikizika kumeneku nthawi zambiri kumatchedwa mawonekedwe apamwamba achiwiri.Pafupifupi zida zonse zapamwamba zimatha kufotokozedwa ndi mitundu iyi yopindika, komanso mitundu yawo yophatikizika, motero amatchedwanso mayunitsi opinda.
3. CAP: cyclic adenosine monophosphate (cAMP) receptor protein CRP (cAMP receptor protein), zovuta zomwe zimapangidwa pambuyo pa kuphatikiza kwa cAMP ndi CRP zimatchedwa activating protein CAP (cAMP activated protein)
4. Palindromic sequence: Mtsinje wowonjezera wotsatizana wa gawo la chidutswa cha DNA, nthawi zambiri malo oletsa ma enzyme.
5. micRNA: Zowonjezera zosokoneza RNA kapena antisense RNA, zomwe zimagwirizana ndi ndondomeko ya mRNA ndipo zingalepheretse kumasulira kwa mRNA.
6. Ribozyme: RNA yokhala ndi ntchito yothandizira, yomwe imagwira ntchito yodziyimira pawokha pakupanga kwa RNA.
7. Motif: Pali madera ena am'deralo omwe ali ndi mawonekedwe ofanana ndi atatu-dimensional ndi topology mumpangidwe wa malo a mamolekyu a protein.
8. Chizindikiro cha peptide: peptide yokhala ndi 15-36 amino acid zotsalira pa N-terminus panthawi ya mapuloteni, omwe amatsogolera transmembrane ya mapuloteni.
9. Attenuator: Kutsatizana kwa ma nucleotide pakati pa chigawo cha opareshoni ndi jini yokhazikika yomwe imathetsa kusindikiza.
10. Malo Amatsenga: Pamene mabakiteriya akukula ndikukumana ndi kusowa kwathunthu kwa amino acid, mabakiteriya adzatulutsa mwadzidzidzi kuti asiye kufotokoza kwa majini onse.Zizindikiro zomwe zimapanga yankho ladzidzidzi ndi guanosine tetraphosphate (ppGpp) ndi guanosine pentaphosphate (pppGpp).Udindo wa PpGpp ndi pppGpp sikuti ndi opereni imodzi kapena yochepa, koma imakhudza chiwerengero chachikulu cha iwo, choncho amatchedwa super-regulators kapena malo amatsenga.
11. Kumtunda kwa promotor element: imatanthawuza kutsatizana kwa DNA komwe kumagwira ntchito yolamulira pazochitika za olimbikitsa, monga TATA m'chigawo cha -10, TGACA m'chigawo cha -35, zowonjezera, ndi zochepetsera.
12. Kufufuza kwa DNA: gawo lolembedwa la DNA lodziwika bwino, lomwe limagwiritsidwa ntchito kwambiri pozindikira masanjidwe osadziwika ndi majini omwe amawonekera pazenera.
13. SD sequence: Ndilo ndondomeko yomangiriza ya ribosome ndi mRNA, yomwe imayendetsa kumasulira.
14. Monoclonal Antibody: Antibody yomwe imagwira ntchito motsutsana ndi antijeni imodzi yokha.
15. Cosmid: Ndi ventor ya DNA yopangidwa mwaluso yomwe imasunga zigawo za COS kumapeto onse a phage ndipo imalumikizidwa ku plasmid.
16. Kuwunika kwa malo a buluu-woyera: LacZ jini (encoding β-galactosidase), enzyme ikhoza kuwononga chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kuti ipange buluu, motero kumapangitsa kuti mtunduwo ukhale buluu.DNA yakunja ikalowetsedwa, jini ya LacZ silingawonetsedwe, ndipo zovuta zake zimakhala zoyera, kuti ziwone mabakiteriya ophatikizanso.Izi zimatchedwa blue-white screening.
17. Cis-acting element: Ndondomeko yeniyeni ya maziko mu DNA yomwe imayang'anira mafotokozedwe a majini.
18. Klenow enzyme: Chidutswa chachikulu cha DNA polymerase I, kupatulapo kuti 5' 3' exonuclease ntchito imachotsedwa mu DNA polymerase I holoenzyme.
19. Anchored PCR: amagwiritsidwa ntchito kukulitsa DNA ya chidwi ndi ndondomeko yodziwika pamapeto amodzi.Mchira wa poly-dG unawonjezedwa kumapeto kwa mndandanda wosadziwika, ndiyeno poly-dC ndi ndondomeko yodziwika idagwiritsidwa ntchito ngati zoyambira pakukulitsa kwa PCR.
20. Puloteni yophatikizika: Jini la puloteni ya eukaryotic limalumikizidwa ndi jini yakunja, ndipo mapuloteni opangidwa ndi kumasulira kwa mapuloteni a jini ndi mapuloteni akunja amawonetsedwa nthawi yomweyo.

Mawu ena a molecular biology

1. Mapu akuthupi a DNA ndi dongosolo limene (restriction endonuclease-digested) zidutswa za molekyulu ya DNA zimasanjidwa.
2. The cleavage ya RNase imagawidwa mu mitundu iwiri (autocatalysis) ndi (heterocatalysis).
3. Pali zinthu zitatu zoyambira mu prokaryotes ndi (IF-1), (IF-2) ndi (IF-3).
4. Mapuloteni a Transmembrane amafuna chitsogozo (signal peptides), ndipo ntchito ya protein chaperones ndi (imathandizira kupindika unyolo wa peptide kuti ugwirizane ndi puloteni).
5. Zinthu zomwe zili mu olimbikitsa zitha kugawidwa m'mitundu iwiri: (zolimbikitsa zoyambira) ndi (zolimbikitsa kumtunda).
6. Zofufuza za biology ya mamolekyulu makamaka zikuphatikizapo magawo atatu: (structural molecular biology), (gene expression and regulation), ndi (DNA recombination technology).
7. Zoyesera ziwiri zazikulu zomwe zikuwonetsa kuti DNA ndi chibadwa ndi (pneumococcus matenda a mbewa) ndi (T2 phage matenda a Escherichia coli).mwayi).
8. Pali kusiyana kwakukulu kuwiri pakati pa hnRNA ndi mRNA: (hnRNA imagawidwa mu njira yosinthira mRNA), (mapeto a 5' a mRNA amawonjezeredwa ndi kapu ya m7pGppp, ndipo pali polyadenylation yowonjezera pa 3 'mapeto a mRNA acid (polyA) mchira).
9. Ubwino wamitundu yambiri ya mapuloteni ndi (subunit ndi njira yachuma yogwiritsira ntchito DNA), (ikhoza kuchepetsa zotsatira za zolakwika zachisawawa mu kaphatikizidwe ka mapuloteni pa ntchito ya mapuloteni), (ntchito ikhoza kukhala yogwira mtima kwambiri komanso mofulumira imatsegulidwa ndi kutsekedwa).
10. Zomwe zili m'mapuloteni opangira mapuloteni oyamba nucleation theory imaphatikizapo (nucleation), (structural enrichment), (final rearrangement).
11. Galactose imakhala ndi mphamvu ziwiri pa mabakiteriya;mbali imodzi (ikhoza kugwiritsidwa ntchito ngati gwero la carbon pa kukula kwa maselo);kumbali ina (ndi gawo la khoma la selo).Chifukwa chake, wothandizira wodziyimira pawokha wa cAMP-CRP S2 amafunikira kuti pakhale kaphatikizidwe kokhazikika pamlingo wakumbuyo;panthawi imodzimodziyo, wothandizira wodalirika wa cAMP-CRP S1 akufunika kuti athetse kaphatikizidwe kapamwamba.Kusindikiza kumayambira ( S2 ) ndi G ndi ( S1 ) popanda G.
12. Ukadaulo wa Recombinant DNA umatchedwanso (gene cloning) kapena (molecular cloning).Cholinga chachikulu ndi (kusamutsa chidziwitso cha majini cha DNA cha chamoyo chimodzi kupita ku chamoyo china).Kuyesera kofananira kwa DNA nthawi zambiri kumakhala ndi njira zotsatirazi: (1) Chotsani jini yomwe mukufuna (kapena jini yakunja) ya chamoyo chopereka, ndikulumikiza ndi molekyulu ina ya DNA (cloning vector) kuti mupange molekyulu ya DNA yatsopano.② The recombinant DNA molekyulu imasamutsidwa mu selo wolandira ndi kubwerezedwa mu selo wolandira.Njira imeneyi imatchedwa kusintha.③ Jambulani ndikuzindikira ma cell omwe alandila omwe adatenganso DNA.④Limbitsani ma cell omwe ali ndi DNA yophatikizanso mochulukira kuti muwone ngati jini yothandizira yakunja ikuwonetsedwa.
13. Pali mitundu iwiri ya kubwereza kwa plasmid: yomwe imayendetsedwa mosamalitsa ndi kaphatikizidwe ka mapuloteni a cell imatchedwa (ma plasmids olimba), ndipo omwe samayendetsedwa mosamalitsa ndi kaphatikizidwe ka mapuloteni a cell amatchedwa (ma plasmids omasuka).
14. Njira ya PCR iyenera kukhala ndi izi: a.Ma DNA oyambira (pafupifupi maziko 20) okhala ndi mindandanda yotsatizana kumapeto kulikonse kwa zingwe ziwiri za jini yomwe mukufuna kusiya.b.Ma enzyme okhala ndi kukhazikika kwamafuta monga: TagDNA polymerase.c, dNTPd, DNA mndandanda wa chidwi monga template
15. Njira yoyambira ya PCR imaphatikizapo magawo atatu: (denaturation), (annealing), ndi (extension).
16. Njira zoyambira za nyama zosasinthika nthawi zambiri zimaphatikizapo: ①Kuyambitsa jini yakunja yopangidwa ndi nyukiliyasi ya dzira lobadwa kapena tsinde la embryonic;②Kuika dzira lopangidwa ndi umuna kapena tsinde la embryonic kulowa m'chiberekero cha mkazi;③Kukula kwathunthu ndi kukula kwa ana omwe ali ndi majini akunja;④ Gwiritsani ntchito nyamazi zomwe zimatha kupanga mapuloteni akunja ngati zoweta kuti zibereke mizere yatsopano ya homozygous.
17. Maselo a Hybridoma amapangidwa ndi hybridizing (spleen B) maselo omwe ali ndi (myeloma) maselo, ndipo popeza (maselo a spleen) amatha kugwiritsa ntchito hypoxanthine ndi (maselo a mafupa) amapereka ntchito zogawanitsa maselo, akhoza kukula mu HAT medium.kukula.
18. Ndi kuzama kwa kafukufuku, mbadwo woyamba wa ma antibodies amatchedwa (polyclonal antibodies), mbadwo wachiwiri (ma antibodies monoclonal), ndi mbadwo wachitatu (ma genetic engineering antibodies).
19. Pakalipano, chibadwa cha tizilombo toyambitsa matenda timayang'ana kwambiri pa baculovirus, zomwe zimawonekera poyambitsa (jini la poizoni wa exogenous);(majini amene amasokoneza moyo wabwinobwino wa tizilombo);(kusintha kwa ma virus).
20. Zomwe zimapangidwira mapuloteni zomwe zimagwirizana ndi zinthu zodziwika bwino za TATA, GC, ndi CAAT mu mammalian RNA polymerase II olimbikitsa ndi (TFIID), (SP-1) ndi (CTF / NF1), motsatira.
makumi awiri ndi mphambu imodzi.Zolemba zoyambirira za RNA polymerase Ⅱ ndi, TFⅡ-A, TFⅡ-B, TFII-D, TFⅡ-E, ndipo mndandanda wawo womangirira ndi: (D, A, B, E).Momwe ntchito ya TFII-D ili (yomangiriza ku bokosi la TATA).
makumi awiri ndimphambu ziwiri.Zambiri mwazinthu zolembera zomwe zimamangiriza ku DNA zimagwira ntchito ngati dimers.Magawo ogwira ntchito a zinthu zolembera zomwe zimamangiriza ku DNA nthawi zambiri zimakhala zotsatirazi (helix-turn-helix), (zinc finger motif), (basic-leucine) zipper motif).
makumi awiri ndi mphambu zitatu.Pali mitundu itatu yoletsa ma endonuclease cleavage modes: (dulani pa 5' mbali ya symmetry axis kuti mupange 5' zomata), (dulani mbali ya 3' ya symmetry axis kuti mupange 3' zomata (kudula pa symmetry axis kuti mupange magawo athyathyathya) ).
makumi awiri ndi mphambu zinayi.Plasmid DNA ili ndi masinthidwe atatu osiyanasiyana: (SC configuration), (oc configuration), (L configuration).Yoyamba mu electrophoresis ndi (SC kasinthidwe).
25. Kachitidwe ka jini yachilendo, makamaka (Escherichia coli), (Yeast), (Tizilombo) ndi (tebulo la maselo a Mammalian).
26. Njira zomwe zimagwiritsidwa ntchito kwambiri pa zinyama zowonongeka ndi: (njira ya kachilombo ka retroviral), (DNA microinjection njira), (njira ya embryonic stem cell).

Kugwiritsa Ntchito Molecular Biology

1. Tchulani ntchito za ma RNA opitilira 5?
Transfer RNA tRNA Transfer amino acid Ribosome RNA rRNA Ribosome constitutes messenger RNA mRNA Protein synthesis template Heterogeneous nuclear RNA hnRNA Precursor of okhwima mRNA yaing'ono nyukiliya RNA snRNA Yophatikizidwa mu hnRNA splicing ndi RNA synthesis yaing'ono RNAs-RNA pulasitiki-RNA yosakanikirana ndi pulasitiki kuzindikira chizindikiro zigawo za thupi Antisense RNA anRNA/micRNA Imawongolera jini Ribozime RNA Enzymatic yogwira RNA
2. Kodi pali kusiyana kotani pakati pa olimbikitsa prokaryotic ndi eukaryotic?
Prokaryotic TTGACA --- TATAAT------ Site Yoyambira-35 -10 Eukaryotic Enhancer---GC ---CAAT----TATAA-5mGpp-Initiation Site-110 -70 -25
3. Ndi mbali ziti zazikulu za kupanga kupanga kwa plasmid zachilengedwe?
Ma plasmids achilengedwe nthawi zambiri amakhala ndi zolakwika, motero sali oyenera kugwiritsidwa ntchito ngati zonyamulira zama genetic engineering, ndipo amayenera kusinthidwa ndikumangidwa: a.Onjezani majini osankhidwa oyenera, monga awiri kapena kuposerapo, omwe ndi osavuta kugwiritsa ntchito posankha, nthawi zambiri ma gene opha maantibayotiki.b.Wonjezerani kapena chepetsani malo oyenera kudula ma enzyme kuti muthandizire kuyanjananso.c.Kufupikitsa kutalika, kudula zidutswa zosafunikira, onjezerani mphamvu zogulira ndikuwonjezera mphamvu yotsegula.d.Sinthani zofananira, kuchokera zolimba kupita zotayirira, kuchokera ku zochepa kukhala zochulukirapo.e.Onjezani zinthu zapadera za majini molingana ndi zofunikira zapadera za uinjiniya wa majini
4. Perekani chitsanzo cha njira yowunikira mosiyanasiyana cDNA yeniyeni?
Maselo awiri a maselo amakonzedwa, jini yomwe imayang'aniridwa imasonyezedwa kapena imasonyezedwa kwambiri mu imodzi mwa maselo, ndipo jini yomwe ikukhudzidwayo siinafotokozedwe kapena kufotokozedwa motsika mu selo lina, ndiyeno jini yomwe ikuyang'aniridwa imapezeka mwa kusakanizidwa ndi kuyerekezera.Mwachitsanzo, pakuchitika ndi kukula kwa zotupa, maselo otupa amawonetsa ma mRNA okhala ndi mawonekedwe osiyanasiyana kuposa maselo abwinobwino.Chifukwa chake, majini okhudzana ndi chotupa amatha kuyang'aniridwa ndi ma hybridization osiyanasiyana.Njira yopangira induction ingagwiritsidwenso ntchito powunikira majini omwe mawu ake amapangidwa.
5. Kubadwa ndi kuwunika kwa mizere yama cell a hybridoma?
Ma cell a spleen B + myeloma cell, onjezerani polyethylene glycol (PEG) kuti alimbikitse kuphatikizika kwa ma cell, ndipo ma cell a splenic B-myeloma fusion omwe amakula mu HAT sing'anga (yomwe ili ndi hypoxanthine, aminopterin, T) ikupitiliza kukulitsa chakudya.Kuphatikizika kwa ma cell kumakhala ndi: ma cell a spleen-spleen fusion: osatha kukula, ma cell a ndulu sangathe kukulitsidwa mu vitro.Maselo ophatikiza mafupa: sangathe kugwiritsa ntchito hypoxanthine, koma amatha kupanga purine kudzera munjira yachiwiri pogwiritsa ntchito folate reductase.Aminopterin amalepheretsa folate reductase ndipo motero sangathe kukula.Maselo ophatikizana ndi mafupa: amatha kukula mu HAT, ma cell a ndulu amatha kugwiritsa ntchito hypoxanthine, ndipo ma cell a mafupa amapereka ntchito yogawa ma cell.
6. Kodi mfundo ndi njira yodziŵira kapangidwe kake ka DNA ndi chiyani ndi njira ya didooxy terminal termination (njira ya Sanger)?
Mfundo yake ndi yogwiritsira ntchito nucleotide chain terminator—2,,3,-dideoxynucleotide kuti athetse kufalikira kwa DNA.Popeza ilibe 3-OH yofunikira kuti apange ma bond 3/5/phosphodiester, ataphatikizidwa mu unyolo wa DNA, unyolo wa DNA sungathe kupitilira.Malinga ndi mfundo ya pairing m'munsi, nthawi iliyonse DNA polymerase imafuna dNMP kuti itenge nawo mbali mu unyolo wa DNA wotambasulidwa, pali zotheka ziwiri, imodzi ndi kutenga nawo mbali mu ddNTP, zomwe zimapangitsa kuti kutha kwa unyolo wa deoxynucleotide;china ndi kutenga nawo mbali mu dNTP , kotero kuti tcheni cha DNA chikhoza kupitilirabe mpaka ddNTP yotsatira idzaphatikizidwa.Malingana ndi njirayi, gulu la zidutswa za DNA zautali wosiyana zomwe zimathera mu ddNTP zikhoza kupezeka.Njirayi ndikugawa m'magulu anayi motsatana ddAMP, ddGMP, ddCMP, ndi ddTMP.Zitachitika, polyacrylamide gel electrophoresis amatha kuwerenga DNA motsatana ndi magulu osambira.
7. Kodi zotsatira zabwino za activator protein (CAP) ndi zotani pa transcript?
Cyclic adenylate (cAMP) receptor protein CRP (cAMP receptor protein), zovuta zopangidwa ndi kuphatikiza kwa cAMP ndi CRP zimatchedwa CAP (cAMPactivated protein).E. coli ikakula m'njira yosowa shuga, kaphatikizidwe ka CAP kumawonjezeka, ndipo CAP imakhala ndi ntchito yoyambitsa zolimbikitsa monga lactose (Lac).Otsatsa ena omwe amadalira CRP alibe mawonekedwe amtundu wa -35 (TTGACA) omwe otsatsa wamba amakhala nawo.Chifukwa chake, ndizovuta kuti RNA polymerase imangirire nayo.Kukhalapo kwa CAP (ntchito): kumatha kusintha kwambiri kukhazikika kwa ma enzyme ndi othandizira.Imawonetsa magawo awiri otsatirawa: ① CAP imathandizira molekyulu ya enzyme kuti isunthike bwino posintha mawonekedwe a olimbikitsa komanso kuyanjana ndi puloteniyo, kuti iphatikize ndi chigawo cha -10 ndikugwira ntchito yolowa m'malo mwa chigawo cha -35.②CAP imathanso kuletsa kumangiriza kwa RNA polymerase kumasamba ena a DNA, potero kumawonjezera mwayi womanga kwa omwe amawalimbikitsa.
8. Kodi ndi zinthu ziti zomwe nthawi zambiri zimaphatikizidwa poyesera kugwirizananso kwa DNA?
a.Chotsani chandamale jini (kapena exogenous jini) wa chamoyo wopereka, ndi enzymatically kulumikiza ndi molekyulu DNA (cloning vekitala) kupanga latsopano recombinant DNA molekyulu.b.Tumizani molekyu ya DNA yophatikizanso mu selo yolandila ndikubwereza ndikuyisunga mu selo yolandila.Njira imeneyi imatchedwa kusintha.c.Jambulani ndikuzindikira maselo omwe alandila omwe adatenga DNA yophatikizanso.d.Misa chikhalidwe ma cell okhala ndi recombinant DNA kuti azindikire ngati jini yothandizira yakunja ikuwonetsedwa.
9. Kupanga laibulale ya majini Njira zitatu zowunikira zophatikizanso zaperekedwa ndipo ndondomekoyi ikufotokozedwa mwachidule.
Kupimidwa kwa maantibayotiki kukana, kulowetsa kukana kukana, kuyang'ana mawanga a buluu-woyera kapena PCR, kuyang'ana mosiyanitsa, DNA probe Ma vector ambiri opangidwa ndi ma cloning amakhala ndi majini olimbana ndi maantibayotiki (anti-ampicillin, tetracycline).plasmid ikasamutsidwa ku Escherichia coli, mabakiteriya amatha kukana, ndipo omwe alibe kusamutsidwa sangakane.Koma silingathe kusiyanitsa ngati linakonzedwanso kapena ayi.Mu vekitala yomwe ili ndi majini awiri osamva, ngati chidutswa cha DNA chachilendo chalowetsedwa mu imodzi mwa jini ndikupangitsa kuti jiniyi ikhale yosagwira ntchito, zowongolera ziwiri zomwe zili ndi mankhwala osiyanasiyana zitha kugwiritsidwa ntchito kuwunika zophatikizanso zabwino.Mwachitsanzo, pUC plasmid ili ndi jini ya LacZ (encoding β-galactosidase), yomwe imatha kuwononga chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kuti ipange buluu, motero kutembenuza mtundu wa buluu.DNA yakunja ikalowetsedwa, jini ya LacZ silingawonetsedwe, ndipo zovuta zake zimakhala zoyera, kuti ziwonetsere mabakiteriya ophatikizanso.
10. Fotokozani njira yopezera nyama zosasinthika kudzera m'maselo a embryonic stem cell?
Maselo a Embryonic stem cell (ES) ndi maselo a embryonic panthawi ya chitukuko cha embryonic, omwe amatha kupangidwa mwaluso komanso kuchulukitsidwa ndikukhala ndi ntchito yosiyana ndi mitundu ina ya maselo.Chikhalidwe cha maselo a ES: Maselo amkati a blastocyst ndi olekanitsidwa komanso otukuka.ES ikakulitsidwa mugawo lopanda chakudya, imasiyana m'maselo osiyanasiyana ogwira ntchito monga ma cell a minofu ndi ma N cell.Ikakulitsidwa mu sing'anga yomwe ili ndi ma fibroblasts, ES imasunga ntchito yosiyanitsa.ES ikhoza kusinthidwa mwachibadwa, ndipo ntchito yake yosiyanitsa ikhoza kuphatikizidwa popanda kusokoneza ntchito yake yosiyanitsa, yomwe imathetsa vuto la kusakanikirana kosasintha.Bweretsani majini achilendo m'maselo a embryonic tsinde, kenaka ikani m'chiberekero cha mbewa zazikazi zapakati, kukhala ana, ndi kuwoloka kuti mupeze mbewa za homozygous.