Tsananguro yematemu eiyo basic molecular biology
1. cDNA uye cccDNA: cDNA iDNA ine tambo mbiri yakagadzirwa ne reverse transcriptase kubva ku mRNA;cccDNA iplasmid yakapetwa kaviri yakavharwa denderedzwa DNA isina kubva kuchromosome.
2. Standard folding unit: protein secondary structure unit α-helix uye β-sheet inogona kuumba mabhuroki ekugadzirisa ane maitiro anokosha ejometri kuburikidza nemhando dzakasiyana-siyana dzinobatanidza polypeptides.Iyi mhando yekupeta yakatemwa inowanzodaidzwa kunzi super secondary chimiro.Anenge ese epamusoro zvimiro anogona kutsanangurwa neaya marudzi ekupeta, uye kunyangwe iwo akasanganiswa marudzi, saka anonziwo akajairwa kupeta zvikamu.
3. CAP: cyclic adenosine monophosphate (cAMP) receptor protein CRP (cAMP receptor protein), yakaoma yakaumbwa mushure mekusanganiswa kwekampu uye CRP inonzi activating protein CAP (cAMP activated protein)
4. Palindromic sequence: The reverse complementary sequement yechikamu cheDNA fragment, kazhinji inorambidza enzyme site.
5. micRNA: Complementary interferring RNA kana antisense RNA, iyo inopindirana nemRNA sequence uye inogona kudzivisa kushandurwa kwemRNA.
6. Ribozyme: RNA ine catalytic chiitiko, iyo inotamba autocatalytic basa mu splicing process yeRNA.
7. Motifi: Kune mamwe matunhu emuno ane akafanana mativi matatu-dimensional chimiro uye topology muchimiro chepakati chemapuroteni mamorekuru.
8. Signal peptide: peptide ine 15-36 amino acid yakasara paN-terminus panguva yeprotein synthesis, iyo inotungamirira transmembrane yeprotein.
9. Attenuator: Kutevedzana kwenucleotide pakati penzvimbo yevashandisi uye geni rechimiro rinomisa kunyorwa.
10. Magic Spot: Apo mabhakitiriya anokura uye anosangana nekushayikwa kwakakwana kweamino acids, mabhakitiriya achabudisa mhinduro yekukurumidzira kumisa kutaura kwemajini ose.Masaini anogadzira iyi mhinduro yechimbichimbi guanosine tetraphosphate (ppGpp) uye guanosine pentaphosphate (pppGpp).Basa rePpGpp uye pppGpp harisi rimwe chete kana mashoma opera, asi rinokanganisa huwandu hukuru hwadzo, saka dzinonzi super-regulators kana mashiripiti mavara.
11. Upstream promoter element: inoreva kutevedzana kweDNA iyo inobata basa rekutonga mubasa rekusimudzira, seTATA munharaunda ye -10, TGACA munharaunda -35, zvinowedzera, uye attenuators.
12. DNA probe: chikamu chakanyorwa cheDNA chine nhevedzano inozivikanwa, iyo inoshandiswa zvakanyanya kuona kutevedzana kusingazivikanwe uye magene ekutarisisa skrini.
13. SD kutevedzana: Ndiyo inosunga nhevedzano ye ribosome uye mRNA, iyo inodzora kududzira.
14. Masoja ekudzivirira chirwere anonzi Monoclonal Antibody: Masoja ekudzivirira chirwere anorwisa antijeni imwe chete.
15. Cosmid: Iyo inyanzvi yakagadzirwa exogenous DNA vector inochengeta matunhu eCOS kumagumo ese ephage uye yakabatana neplasmid.
16. Blue-white spot screening: LacZ gene (encoding β-galactosidase), iyo enzyme inogona kuparadza chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kuti ibudise bhuruu, nokudaro ichiita kuti mhirizhonga yebhuruu.Kana iyo exogenous DNA yaiswa, iyo LacZ gene haigone kuratidzwa, uye dhizaini ichena, kuitira kuti utarise mabhakitiriya anosanganisirwa.Izvi zvinonzi blue-white screening.
17. Cis-acting element: Kutevedzana kwakananga kwezvigadziko muDNA zvinogadzirisa gene expression.
18. Klenow enzyme: Chimedu chikuru cheDNA polymerase I, kunze kwekuti 5' 3' exonuclease basa inobviswa muDNA polymerase I holoenzyme.
19. Anchored PCR: yakashandiswa kukwidziridza DNA yekufarira nekutevedzana kunozivikanwa pane imwe mugumo.Muswe wepoly-dG wakawedzerwa kune rumwe rutivi rwekutevedzana kusingazivikanwe, uye ipapo iyo poly-dC uye iyo inozivikanwa kutevedzana yakashandiswa semaprimers ePCR amplification.
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Mamwe mazwi emolecular biology
1. Mepu yemuviri yeDNA ndiyo marongero ayo (restriction endonuclease-digested) zvidimbu zvemorekuru weDNA zvinorongwa.
2. Kutsemuka kweRNase kwakakamurwa kuva marudzi maviri (autocatalysis) uye (heterocatalysis).
3. Pane zvinhu zvitatu zvokutanga muprokaryotes ndezvi (IF-1), (IF-2) uye (IF-3).
4. Mapuroteni eTransmembrane anoda kutungamirirwa (signal peptides), uye basa reprotein chaperones (inobatsira kupeta peptide chain mukugadzirwa kwekuzvarwa kweprotein).
5. Zvinhu zviri mumapromota zvinogona kukamurwa kuita mhando mbiri: (core promoter elements) uye (upstream promotioner elements).
6. Zvinyorwa zvekutsvakurudza zvemolecular biology zvinonyanya kusanganisira zvikamu zvitatu: (structural molecular biology), (gene expression and regulation), uye (DNA recombination technology).
7. Zviedzo zviviri zvakakosha zvinoratidza kuti DNA ndiyo genetic material ndeye (pneumococcus infection yemakonzo) uye (T2 phage infection yeEscherichia coli).zvinogona).
8. Pane misiyano miviri mikuru pakati pehnRNA uye mRNA: (hnRNA inoparadzaniswa mukugadzirisa kuchinjwa kuva mRNA), (iyo 5 'kuguma kwemRNA inowedzerwa ne m7pGppp cap, uye kune imwe polyadenylation pa 3' kuguma kwemRNA acid (polyA) muswe).
9. Zvakanakira zve-multi-subunit fomu yeprotein ndeye (subunit inzira yehupfumi yekushandiswa kweDNA), (inogona kuderedza kukanganisa kwekukanganisa muprotein synthesis pamabasa eprotein), (basa rinogona kunyatsoshanda uye nekukurumidza rinovhurwa uye rakavharwa).
10. Nyaya huru yeprotein folding mechanism yekutanga nucleation theory inosanganisira (nucleation), (kugadziriswa kwemaitiro), (rearrangement yekupedzisira).
11. Galactose ine maitiro maviri pabhakitiriya;kune rumwe rutivi (inogona kushandiswa se carbon source yekukura kwesero);kune rumwe rutivi (iriwo chikamu chesero rusvingo).Naizvozvo, cAMP-CRP-yakazvimiririra inosimudzira S2 inodiwa kune yekusingaperi synthesis padanho rekumashure;panguva imwecheteyo, cAMP-CRP-inotenderera inosimudzira S1 inodiwa kuti igadzirise iyo yepamusoro-yepamusoro synthesis.Kunyorerwa kunotanga kubva ( S2 ) naG uye kubva ( S1 ) isina G.
12. Recombinant DNA teknolojia inozivikanwawo se (gene cloning) kana (molecular cloning).Chinangwa chikuru ndechekuti (kuendesa ruzivo rwemajini DNA mune chimwe chipenyu kuenda kune chimwe chipenyu).Kuedza kusanganisazve kweDNA kunowanzo sanganisira nhanho dzinotevera: (1) Bvisa jini (kana gene exogenous) yechipenyu cheanopa, woibatanidza zvine enzymatic kune imwe molecule yeDNA (cloning vector) kuti igadzire molecule nyowani reDNA.② Iyo recombinant DNA molecule inotamirwa musero reanogamuchira uye inodzokororwa musero rinogamuchira.Iyi nzira inonzi shanduko.③ Vhura uye tsvaga iwo maseru anogashira atora iyo recombinant DNA.④Gadzira maseru ane recombinant DNA muhuwandu hukuru kuti uone kana gene rerubatsiro rekunze rinoratidzwa.
13. Kune marudzi maviri ekudzokorora kweplasmid: iyo inonyatsodzorwa nehost cell protein synthesis inonzi (tight plasmids), uye iyo isina kunyatsodzorwa nehost cell protein synthesis inonzi (relaxed plasmids).
14. PCR reaction system inofanira kuva nemamiriro anotevera: a.DNA primers (mabhesi angangoita makumi maviri) ane anowirirana akatevedzana kumagumo ega ega tambo mbiri dzejini rinoda kuparadzaniswa.b.Enzymes ine kugadzikana kwekupisa senge: TagDNA polymerase.c, dNTPd, DNA kutevedzana kwekufarira se template
15. Maitiro ekutanga maitiro ePCR anosanganisira matanho matatu: (denaturation), (annealing), uye (kuwedzera).
16. Maitiro ekutanga emhuka dzakashanduka kazhinji anosanganisira: ①Kuunzwa kwegene rekunze rakagadzirwa mukati mezai rakasangana kana embryonic stem cell;②Kusimwa kwezai rakaiswa fetiraiza kana embryonic stem cell muchibereko chemukadzi;③Kukura uye kukura kwe embryonic Kune vana vane majini ekunze;④ Shandisa mhuka idzi dzinogona kuburitsa mapuroteni ekunze semombe yekuberekesa kubereka mitsetse mitsva yehomozygous.
17. Hybridoma cell lines inogadzirwa ne hybridizing (spleen B) masero ane (myeloma) masero, uye sezvo (spleen masero) anogona kushandisa hypoxanthine uye (mapfupa emapfupa) anopa mabasa ekuparadzanisa masero, anogona kukura muHAT yepakati.kukura.
18. Nekudzika kwekutsvakurudza, chizvarwa chekutanga chezvirwere zvinonzi (polyclonal antibodies), chizvarwa chechipiri (monoclonal antibodies), uye chizvarwa chechitatu (genetic engineering antibodies).
19. Parizvino, genetic engineering yezvipembenene zvezvipembenene zvinonyanya kutarisa baculovirus, iyo inoratidzwa mukuiswa kwe (exogenous toxin gene);(majini anovhiringidza mararamiro akajairika ezvipembenene);(kugadziriswa kwemajini ehutachiona).
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makumi maviri nerimwe.Izvo zvakakosha zvinyorwa zveRNA polymerase Ⅱ ndezvi, TFⅡ-A, TFⅡ-B, TFII-D, TFⅡ-E, uye kutevedzana kwavo kunosunga ndekwe: (D, A, B, E).Imo basa reTFII-D riri (kusunga kuTATA bhokisi).
makumi maviri nemaviri.Zvizhinji zvezvinyorwa zvekunyora zvinosunga kuDNA zvinoshanda nenzira ye dimers.Nzvimbo dzinoshanda dzezvinyorwa zvinyorwa zvinosunga kuDNA zvinowanzotevera (helix-turn-helix), (zinc finger motif), (basic-leucine) zipper motif).
makumi maviri nenhatu.Kune marudzi matatu ekurambidza endonuclease cleavage modes: (cheka pa5' side re symmetry axis kugadzira 5' inonamira magumo), (cheka pa 3' side re symmetry axis kugadzira 3' inonamira magumo (cheka pa symmetry axis kugadzira zvikamu zvakati sandara) ).
makumi maviri nemana.Plasmid DNA ine zvigadziriso zvitatu zvakasiyana: (SC configuration), (oc configuration), (L configuration).Yekutanga mu electrophoresis ndeye (SC configuration).
25. Exogenous gene expression systems, kunyanya (Escherichia coli), (Yeast), (Chipembenene) uye (Mammalian cell table).
26. Nzira dzinowanzoshandiswa dzemhuka dze transgenic ndeidzi: (retroviral infection method), (DNA microinjection method), (embryonic stem cell method).
Kushandisa Molecular biology
1. Doma mabasa anopfuura mashanu eRNA?
Shandura RNA tRNA Shandura amino acid Ribosome RNA rRNA Ribosome inoumba mutumwa RNA mRNA Protein synthesis template Heterogeneous nuclear yenyu RNA hnRNA Precursor of mature mRNA nyukireya diki RNA snRNA Inobatanidzwa muhnRNA splicing splicing RNAs splicing RNA splicing RNAs Splicing DikiRNA splicing RNAssl scl cytoplasmic RNA scl sc. chiratidzo chekuziva muviri zvikamu Antisense RNA anRNA/micRNA Inogadzirisa gene kutaura Ribozyme RNA Enzymatically inoshanda RNA
2. Ndeupi musiyano mukuru pakati peprokaryotic uye eukaryotic vanokurudzira?
Prokaryotic TTGACA --- TATAAT------Initiation Site-35 -10 Eukaryotic Enhancer---GC ---CAAT----TATAA-5mGpp-Initiation Site-110 -70 -25
3. Ndezvipi zvinhu zvakakosha zvekuvakwa kwechisikigo cheplasmid chaiyo?
Maplasmids echisikigo anowanzo kuve akaremara, saka haana kukodzera kushandiswa sevatakuri vemajini engineering, uye anofanirwa kugadziridzwa nekuvakwa: a.Wedzera akakodzera majene echiratidzo, akadai maviri kana anopfuura, ayo ari nyore kushandisa pakusarudza, kazhinji majene emishonga inorwisa mabhakitiriya.b.Wedzera kana kuderedza nzvimbo dzakakodzera dzekucheka enzyme kuti dzifambise kubatanidzwa.c.Pfupisa kureba, cheka zvimedu zvisingakoshi, vandudza kupinza kwekunze uye kuwedzera huwandu hwekurodha.d.Shandura replicon, kubva kune yakasimba kuenda kune yakasununguka, kubva kune mashoma makopi kuenda kune mamwe makopi.e.Wedzera zvakakosha genetic zvinhu zvinoenderana neakakosha zvinodiwa zvemajini engineering
4. Ipa muenzaniso wenzira yekutarisa mutsauko wetishu-chaiyo cDNA?
Masero maviri evanhu akagadzirirwa, jini rinotarirwa rinoratidzwa kana rakanyatsoratidzwa mune rimwe masero, uye jini rinotarirwa harina kuratidzwa kana kuderedzwa kuratidzwa mune rimwe sero, uye ipapo jini rinotarirwa rinowanikwa nekusanganiswa uye kuenzanisa.Semuyenzaniso, panguva yekuitika uye kukura kwemamota, masero ebundu achapa mRNAs ane mazinga ekutaura akasiyana pane akajairwa maseru.Naizvozvo, majini ane hukama nebundu anogona kuongororwa nekusiyanisa hybridization.Iyo induction nzira inogona zvakare kushandiswa kuvharidzira kunze kwemajini ane kutaura kunokonzeresa.
5. Chizvarwa uye kuongororwa kwehybridoma cell mitsara?
Spleen B masero + myeloma masero, wedzera polyethylene glycol (PEG) kukurudzira sero fusion, uye splenic B-myeloma fusion masero anokura muHAT medium (ine hypoxanthine, aminopterin, T) inoramba ichiwedzera kudya.Iyo sero fusion ine: spleen-spleen fusion masero: haakwanise kukura, spleen masero haagone kugadzirwa mu vitro.Bone-bone fusion maseru: haigone kushandisa hypoxanthine, asi inogona kugadzira purine kuburikidza neyechipiri nzira uchishandisa folate reductase.Aminopterin inhibits folate reductase uye nokudaro haigoni kukura.Bone-spleen fusion masero: anogona kukura muHAT, spleen masero anogona kushandisa hypoxanthine, uye masero emapfupa anopa sero division basa.
6. Ndeipi musimboti uye nzira yekuziva chimiro chekutanga cheDNA nenzira ye didioxy terminal termination method (Sanger method)?
Nheyo yacho ndeyekushandisa nucleotide chain terminator-2,,3,-dideoxynucleotide kugumisa kuwedzera kweDNA.Sezvo isina 3-OH inodiwa pakuumbwa kwe3 / 5 / phosphodiester zvisungo, kana yakabatanidzwa muketani yeDNA, cheni yeDNA haigoni kuwedzerwa kuwedzerwa.Zvinoenderana nemusimboti we base pairing, pese apo DNA polymerase inoda dNMP kuti itore chikamu mune inowanzowedzerwa DNA cheni, pane zvingave zviviri, imwe ndeyekutora chikamu muddNTP, izvo zvinoguma nekugumiswa kwedeoxynucleotide chain extension;imwe yacho ndeyekutora chikamu mu dNTP , kuitira kuti cheni yeDNA irambe ichiwedzera kusvika ddNTP inotevera yabatanidzwa.Maererano nenzira iyi, boka rezvimedu zveDNA zvehurefu hwakasiyana zvinopera muddNTP zvinogona kuwanikwa.Nzira yacho ndeyekukamura kuita mapoka mana zvakateerana ddAMP, ddGMP, ddCMP, uye ddTMP.Mushure mekuita, polyacrylamide gel electrophoresis inogona kuverenga kutevedzana kweDNA zvichienderana nemabhendi ekushambira.
7. Ndeipi iyo yakanaka regulation mhedzisiro ye activator protein (CAP) pane chinyorwa?
Cyclic adenylate (cAMP) receptor protein CRP (cAMP receptor protein), yakaoma inoumbwa nemusanganiswa wecAMP uye CRP inonzi CAP (cAMPactivated protein).Kana E. coli yarimwa mune yepakati inoshaya glucose, iyo synthesis yeCAP inowedzera, uye CAP ine basa re activate mapromota akadai lactose (Lac).Vamwe vanotsigira CRP-vanotsamira vanoshaya iyo yakajairwa -35 dunhu sequence ficha (TTGACA) iyo yakajairika mapromota.Naizvozvo, zvakaoma kuti RNA polymerase isungirire pairi.Kuvapo kweCAP (basa): inogona kuvandudza zvakanyanya kusungirirwa kwe enzyme uye mutsigiri.Iyo inonyanya kuratidza zvinhu zviviri zvinotevera: ① CAP inobatsira iyo enzyme molecule kutenderera nenzira kwayo nekushandura conformation yeanosimudzira uye kudyidzana neiyo enzyme, kuitira kuti ibatanidze ne -10 dunhu uye kuita basa rekutsiva basa re -35 dunhu.②CAP inogonawo kurambidza kusungwa kweRNA polymerase kune dzimwe nzvimbo muDNA, nekudaro ichiwedzera mukana wekusunga kune chaiwo anosimudzira.
8. Matanho api anowanzobatanidzwa mukuedza kusanganisazve kweDNA?
a.Bvisa jini (kana exogenous gene) yeanopa chipenyu, uye enzymatically uchibatanidza kune imwe DNA molecule (cloning vector) kuti igadzire itsva recombinant DNA molecule.b.Fambisa recombinant DNA molecule musero rinogamuchira uye dzokorora uye chengetedza musero rinogamuchira.Iyi nzira inonzi shanduko.c.Vhura uye tsvaga iwo masero anogamuchira atora iyo recombinant DNA.d.Misa tsika masero ane recombinant DNA kuti aone kana iyo yekunze gene yerubatsiro inoratidzwa.
9. Kuvakwa kweraibhurari yemajini Nzira nhatu dzekutarisisa marecombinants dzinopihwa uye maitiro anotsanangurwa muchidimbu.
Antibiotic resistance screening, insertional inactivation of resistance, blue-white spot screening kana PCR screening, differential screening, DNA probe Mazhinji cloning vectors anotakura ma antibiotic resistance genes (anti-ampicillin, tetracycline).Apo plasmid inotamisirwa muEscherichia coli, mabhakitiriya anowana kupikisa, uye avo vasina kutamiswa havazopikisi.Asi haigoni kusiyanisa kuti yakarongwazve here kana kuti kwete.Muvheta rine majini maviri ekuramba, kana chimedu cheDNA chekune imwe nyika chikaiswa mune rimwe remajini uye chikakonzera kuti jena risashande, zvidzoreso zveplate zviviri zvine mishonga yakasiyana zvinogona kushandiswa kuvheneka kuti zvionekwe kune zvinosanganiswa zvakanaka.Semuenzaniso, pUC plasmid ine LacZ gene (encoding β-galactosidase), iyo inogona kuora chromogenic substrate X-gal (5-bromo-4-chloro-3-indole-β-D-galactoside) kuti ibudise bhuruu, nokudaro kushandura strain blue.Kana iyo yekunze DNA yakaiswa, iyo LacZ gene haigone kuratidzwa, uye dhizaini ichena, kuitira kuti itarise mabhakitiriya anobatana.
10. Tsanangura nzira yekutanga yekuwana mhuka dzakashanduka kuburikidza ne embryonic stem cell?
Embryonic stem cells (ES) ndiwo masero embryonic panguva yekukura kwe embryonic, iyo inogona kugadzirwa nemaitiro uye kuwedzera uye iine basa rekuparadzanisa mune mamwe marudzi emasero.Tsika yemasero eES: Iyo yemukati sero misa ye blastocyst yakaparadzaniswa uye inogadzirwa.Kana ES yakasimwa mune yekudyisa-isina layer, inozosiyanisa kuita akasiyana siyana anoshanda maseru semhasuru maseru uye N masero.Kana yakasimudzwa mune svikiro ine fibroblasts, ES inochengetedza basa rekusiyanisa.ES inogona kugadzirwa nemajini, uye basa rayo rekusiyanisa rinogona kubatanidzwa pasina kukanganisa basa rayo rekusiyanisa, rinogadzirisa dambudziko rekubatanidza kusina.Isai exogenous majini muembryonic stem cells, wozoisa muchibereko chembeva dzechikadzi dzine pamuviri, dzoita pups, uye yambuka kuti uwane homozygous mbeva.
