Hielscher faka'ata tekinolosia

Lysis toʻotoʻo ʻo bl21 ngaahi selo ʻe Ultrasonication

Ko e ngaahi selo BL21 ko ha mafasia ia ʻo e E. kololi ʻoku fakaʻaongaʻi lahi ʻi he fekumi laboratories, biotechnology mo e ngaahi meʻa fakapisinisi koeʻuhi ko ʻenau malava ke fakahaaʻi polotini lelei ʻaupito. Ko e ultrasonic toʻotoʻo, lysis mo e ivi toʻo ʻuli ko e founga angamaheni ia ke fakamavaheʻi mo tanaki ʻa e polotini kuo fakataumuʻa ki ai mei he loto Mopaila ʻo bl21 ngaahi selo. Ultrasonication fakahohaʻasi ʻa e meʻa toʻotoʻo pea tukuange kotoa ʻa e polotini, ʻo maʻu ai ʻa e 100% ʻo e ivi ʻoku maʻu.

BL21 e ngaahi selo ki hono fakahaaʻi ʻo e ivi

E. coli bacteria such as BL21 cells are commonly lysed by ultrasonication in order to release expressed proteins, such as recombinant proteinsKo bl21 toʻotoʻo ko ha faitoʻi taukei E. kololi hangatamaki faingataʻaʻia feʻunga ke liliu mo e tuʻunga maʻolunga ʻo e fakahaaʻi ʻo fakaʻaongaʻi ha T7 RNA polymerase-IPTG induction System. ʻOku fakaʻata ʻe he ngaahi selo BL21 ha ivi feʻunga ke fakahaaʻi ha faʻahinga Sini ʻoku ne puleʻi ha T7 promoter. Ko e E. kololi BL21 (DE3) ko ha T7 RNA polymerase ʻoku ne fakatahaʻi ʻa e ivi ʻoku ne fakatahaʻi mo T7 promoter-based vectors pea ʻoku fakaʻaongaʻi lahi ia ʻi he laboratories mo e faʻa ngaue ke fakatupu recombinant polotini. ʻI BL21 (DE3), ko hono fakahaaʻi ʻo e sini e encoding ʻoku hiki ia ʻe he chromosomally encoded T7 RNA polymerase (T7 RNAP), ʻa ia transcribes tuʻo valu ʻi he angamaheni ko e. kololi RNAP. ʻOku ngaohi ʻe he meʻa ni ʻa e faingataʻaʻia BL21 (DE3) lelei ʻaupito pea liliu ia ki he taha ʻo e ngaahi founga toʻotoʻo ʻoku fie maʻu taha.

Protocol ki he ultrasonic Lysis mo e ivi toʻo hingoa mei he ngaahi selo BL21

Ko e lysis toʻotoʻo ʻo e ngaahi selo BL21 ʻoku meimei ke fakahoko pe ia ʻaki hono fakaʻaongaʻi ʻo e ultrasonication ʻi hono fakatahaʻi mo ʻelemeniti ko lauroyl sarcosinate (ʻoku toe ʻiloa pe ko sarkosyl) ko lysis ngataʻanga. ʻOku loi ʻa e ngaahi lelei ʻo ultrasonic fakahohaʻasi toʻotoʻo mo e ivi toʻo ʻuli ʻi he ala falalaʻanga, reproducibility pea pehe ki he ngaue faingofua, malu mo vave ʻo e ultrasonicators. ʻOku ʻomi ʻe he ngaahi founga fakangaue ʻi lalo ha fakahinohino fakasitepa ki he ultrasonic BL21 toʻotoʻo lysis:

Vakili-fa'ahinga insonifier UP200St ki he lysis

Ultrasonic toʻotoʻo disruptor UP200St mo e S26d2 Micro ki he lysis mo e ivi toʻo hingoa

Kole 'a e fakamatala




Fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


  • Koeʻuhi ke toʻo ʻa e polotini, BL21 hangatamaki pellets naʻe resuspended ʻi he 50 ml ʻo e ʻaisi momoko ʻelemeniti ko Tris-EDTA (STE) (ʻa ia ʻoku ʻi he 10 mM Tris-HCL, pH 8.0, 1 mM EDTA, 150 mM.
  • ʻOku tanaki atu ʻa e, 500 ul ʻo lysozyme (10 mg/ml) pea incubated ʻa e ngaahi selo ʻi he ʻaisi ʻi ha miniti ʻe 15.
  • Hili ia, 500 ul ʻo DTT mo e 7 ml ʻo sarkosyl (10% (w/v) naʻe faʻu ʻi STE ngataʻanga).
  • ʻOku mahuʻinga ke tauhi ʻa e fakamaʻa kotoa buffers ʻaisi-momoko pea ke tauhi ʻa e ngaahi sipinga ʻi he ʻaisi he taimi kotoa pe. ʻOku totonu ke fakahoko ʻa e ngaahi sitepu fakamaʻa kotoa pē ʻi he loki momokó kapau ʻe lava.
  • VialTweeter at the ultrasonic processor UP200ST

  • Ki ultrasonic lysis mo e ivi toʻo ʻuli, ʻoku sonicated ʻa e ngaahi sipinga ʻi he VialTweeter MultiSample Ultrasonicator ki he 4 x 30 Sec ʻi he 100% amplitude mo ha miniti ʻe 2 ʻi he vahaʻa ʻo e sonication takitaha. Pe, ko ha fekumi-type ultrasonic homogenizer mo e Micro-tip e.g., UP200Ht mo e S26d2 (3 x 30 Sec, miniti ʻe 2. kiʻi malolo ʻi he vahaʻa ʻo ultrasonic siakale, 80% amplitude).
  • Ke maʻu ha ngaahi sitepu ki hono fakamaʻa lahi ange, kuo pau ke tauhi ʻa e ngaahi sipinga ʻi he ʻaisi pe pe tauhi ʻi he 80 ° c kae ʻoua kuo toe fai ha ngaue ki ai.

Ultrasonic Lysis ʻi he malumalu ʻo e mafana ʻo e ʻea

ʻOku mahuʻinga ʻa e mapuleʻi totonu mo falalaʻanga ʻo e ʻea ʻi he taimi ʻoku nau tokangaʻi ai ʻa e ngaahi sipinga totonu. ʻOku kamata ʻe he mafana ʻo e ʻea ʻa e thermally ʻoku ne fakaʻaiʻai ʻa e maʻulalo ʻi ha ngaahi sipinga.
ʻI he hoko ʻa e sipinga fakamisini kotoa pe, ʻoku fakatupu ʻe sonication ha vela. Neongo ia, ʻe lava ke mapuleʻi lelei e mafana ʻo e ngaahi sipinga ʻi he taimi ʻoku fakaʻaongaʻi ai e VialTweeter. ʻOku mau ʻoatu ha ngaahi fili kehekehe ke vakaiʻi mo puleʻi ʻaki e mafana hoʻo sipinga lolotonga hono teuteuʻi kinautolu ʻaki e VialTweeter mo e VialPress ke ʻanalaiso.

  1. Vakaiʻi ʻa e sipinga ʻo e ʻea: ko e ultrasonic processor UP200St, ʻa ia ʻoku ne teke ʻa e VialTweeter, ʻoku fakanaunau ʻaki ia ha polokalama fakakomipiuta fakapotopoto mo ha mafana pluggable sensor resistance. Palaki ʻa e mafana ʻo e ʻea sensor resistance ki he UP200St pea fakahu ʻa e tokoni ʻa e ʻea sensor resistance ʻi he taha ʻo e ngaahi sipinga ʻo e tiupi. ʻI he naunau fakaʻilekitulonika coloured, te ke lava ʻo fokotuʻu ʻi he menu ʻo e UP200St ha mafana pau ki hoʻo sipinga sonication. ʻE taʻofi leva ʻa e ultrasonicator ʻi he taimi ʻe aʻu ai ʻa e mafana ʻo e ʻea pea kiʻi malolo kae ʻoua kuo aʻu ʻa e sipinga ʻo e ʻea ki he tuʻunga ʻulungaanga mahuʻinga maʻulalo ange ʻo e mafana ʻo e ʻ∆ mafana. Pea toe kamata leva ʻa e sonication. ʻOku taʻofi ʻe he founga fakapotopoto ko ʻeni ʻa e maʻulalo ʻoku ne fakaʻaiʻai ʻa e ʻea.
  2. ʻE lava ke tomuʻa ʻita ʻa e konga VialTweeter. Faʻo ʻa e konga VialTweeter (ko e sonotrode pe taʻe transducer!) ki he ʻaisi pe ʻaisi ke teuteu ke mokomoko ʻa e konga titanium ʻoku tokoni ke toloi ʻa e mafana ʻo e ʻea ʻi he sipinga. Kapau ʻe lava, ʻe lava ke tomuʻa ʻita foki mo e sipinga.
  3. Fakaʻaongaʻi ʻa e ʻaisi momoa ke mokomoko lolotonga e sonication. Fakaʻaongaʻi ha laulau mamaha ʻoku fonu ʻi he ʻaisi momoa pea tuku ʻa e VialTweeter ʻi he ʻaisi koeʻuhi ke lava ʻo molia vave ʻa e vela.

ʻOku fakaʻaongaʻi ʻe he kau fakatau fakaemamani lahi ʻa e VialTweeter mo e VialPress ki heʻenau sipinga teuteu fakaʻaho ʻi he ngaue totonu, biochemical, fakafaitoʻo mo kiliniki laboratories. Ko e polokalama fakakomipiuta mo e mafana ʻo e ʻea ʻoku ne puleʻi ʻa e UP200St processor, ʻoku reliably mapuleʻi ʻa e mafana mo e sipinga ʻo e ʻea vela ʻoku ne fakaʻehiʻehi mei ai. ʻOku hanga ʻe ultrasonic sipinga ʻo e teuteu mo e VialPress VialPress ʻo fakahaofi ha ngaahi ola falalaʻanga mo reproducible!

Kumi ʻa e tokolahi lelei ultrasonic Disruptor ki hoʻo tohi kole Lysis

Hielscher's industrial processors of the hdT series can be comfortable and user-friendly operated via browser remote control.Ko Hielscher Ultrasonics ko ha kautaha taukei taimi loloa ia ʻo e ngaue maʻolunga ultrasonic disrupters mo homogenizers ki laboratories, ngaahi founga meʻafua ki ʻolunga mo e ngaahi meʻa fakamisini. Ko hoʻo hangatamaki lahi ʻo e anga fakafonua, ko hoʻo fakatotolo pe taumuʻa ʻo e ngaue pea mo e lahi ʻo e meʻa toʻotoʻo ke fai ʻi he houa pe ʻaho, ko ha ngaahi meʻa mahuʻinga ia ke maʻu ai ʻa e ultrasonic toʻotoʻo disruptor ki hoʻo tohi kole.
ʻOku ʻomi ʻe Hielscher Ultrasonics ha ngaahi founga kehekehe ki he hoko sonication ʻo e ngaahi sipinga fakavahaʻa-taʻu (ʻo aʻu ki he 10 mo e VialTweeter) mo ha ngaahi sipinga lahi (hange ko e, microtiter ʻu lauʻi peleti/ʻELISA mo e UIP400MTP), pea pehe ki he loki fakatotolo ʻiloa fekumi ultrasonicator mo e ngaahi tuʻunga malohi kehekehe mei he 50 ki he 400 Watts ki he ngaahi meʻa lalahi ultrasonic processors ʻo aʻu ki he 16,000 ʻi he ʻiuniti ki he fakahohaʻasi fakakomesiale mo e ivi toʻo ʻuli lahi. ʻOku langa ʻa e Hielscher ultrasonicators kotoa pe ki he ngaue 24/7/365 ʻi he kavenga kakato. Ko e Robustness mo e ala falalaʻanga ko e ngaahi tefitoʻi fotunga ia ʻo ʻetau ngaahi meʻangaue ultrasonic.
ʻOku fakanaunau ʻa e ultrasonic fakaʻilekitulonika kotoa pe homogenizers ʻaki ʻa e polokalama fakakomipiuta poto, coloured ala ki he fakaʻaliʻali mo e fakamatala fakahangatonu protocolling, ʻa ia ʻoku ne ʻai ʻa e meʻangaue ultrasonic ki ha meʻangaue ʻoku faingamalie ke fai ʻi he fale fakatotolo fakakemi mo e ngaahi naunau ngaohiʻanga naunau.
Tau ʻilo muʻa, ko e ha ʻa e faʻahinga selo, ko e ha ʻa e voliume, mo e tuʻo lahi pea mo e meʻa ʻoku fakataumuʻa ke ke fakahoko ʻaki hoʻo ngaahi sipinga totonu. Te mau fokotuʻu atu ʻa e ultrasonic toʻotoʻo lelei taha disruptor ki hoʻo ngaahi fie maʻu ki he founga ngaue.

ʻOku ʻoatu ʻe he tepile ʻi lalo ha fakaʻilonga ʻo e fakafuofua ki he tuʻunga ʻo ʻetau ngaue ultrasonic mei he nima homogenizers mo MultiSample Ultrasonicators ki he ngaahi ultrasonic processors ki he ngaahi polokalama fakakomesiale:

Kulupu (batch) 'o e tohi 'Oku tafe mai 'a e 'ea 'Oku fokotu'u atu 'a e ngaahi me'angaue
96-ngaahi lauʻi peleti/microtiter n.a. UIP400MTP
10 à 0.5 ki he 1.5 mL n.a. VialTweeter 'i he UP200St
0.01 ki he 250mL 5 ki he 100mL/miniti UP50H
0.01 ki he 500mL 10 ki he 200mL/miniti 'e UP100H
mL 'i he 10 ki he 2000 20 ki he 400mL/miniti 'e UP200Ht, UP400St
0.1 ki he 20L 0.2 ke 4L/miniti 'e UIP2000hdT
10 ki he 100L 2 ki he 10L/miniti 'e UIP4000hdT
n.a. 10 ki he 100L/miniti 'e UIP16000
n.a. lalahi fakataha'i 'o e UIP16000

Fetu'utaki mai kiate kimautolu! / Kole kiate kitautolu!

Kole ki ha fakamatala lahi ange

Kataki 'o ngaue 'aki e foomu 'i lalo ke kole ha toe fakamatala ki he ultrasonic processors, faka'aonga'i mo e totongi. Te mau fiefia ke alea'i e founga 'oku mo 'i ai mo ho'omou ngaahi fie ma'u ultrasonic he polokalama!









Kataki 'o fakatokanga'i ange 'etau Tu'utu'uni totonu fakatautaha.


Ultrasonicator UP200Ht with microtip S26d2 for ultrasonic lysis of biological samples

ultrasonicator UP200Ht mo e 2mm microtip S26d2 ki he sonication ʻo ha fanga kiʻi sipinga iiki

Ngaahi tohi/fakamo'oni fakafolofola



Ngaahi mo'oni'i me'a 'oku mahu'inga ke 'ilo'i

Escherichia Kololi pekitilia

Ko Escherichia kololi ko ha faʻahinga pekitilia, ʻoku ʻikai ko ha faʻu spore, Gram-kovi pea ʻoku fakafotunga ia ʻe hono fotunga ʻo ha vaʻa ukamea hangatonu. E. kololi ʻoku ʻi he ʻatakai, meʻakai, mo e ngakau ʻo e kau tau mo e fanga monumanu. E. kololi ʻoku faʻa motile ʻaki hono fakaʻaongaʻi ʻo e peritrichous flagella, ka ʻoku ʻi ai foki mo ha faʻahinga momotile. E. kololi ʻoku ui ko e facultatively anaerobic chemoorganotroph organisms, ʻa ia ko hono ʻuhinga ʻoku nau lava ai ʻo manava mo fermentative metabolism. Ko e lahi taha ʻo e ngaahi faʻahinga ʻo e kololi ʻoku angaʻofa mo fakahoko ʻa e ngaahi ngaue ʻaonga ʻi he sino, hange ko hono mapuleʻi ʻo e tupu ʻa e hangatamaki fakafaʻahinga fakatuʻutamaki, synthesising vaitamini mo e ala meʻa pehe.
ʻOku hoko ʻa Escherichia kololi ko e pekitilia ʻo e faʻahinga ko ia ʻoku ui ko e ko ha kulupu makehe ʻo e E. kololi, ʻa ia ʻoku fakaʻaongaʻi lahi ʻi he fekumi ke fakatotoloʻi ngaahi meʻa kehe hange ko bacteriophage ongoʻingofua pe ngaahi founga liliu fakangatangata. ʻIkai ngata ai, ʻoku fakamahuʻingaʻi ʻa e pekitilia ʻa e kololi ko ha kau falalaʻanga ke fakahaaʻi ha ivi ʻi he biotechnology mo e saienisi ʻo e moʻui laboratories. Hange ko ʻeni, ʻoku fakaʻaongaʻi ʻa e E. kololi ke synthesize meʻa koʻeni hange ko polotini mo oligosaccharides ʻi he meʻafua ʻo e ngaahi meʻa fakapisinisi. Koeʻuhi ko ha ngaahi meʻa pau hange ko e protease ʻoku ʻikai ke kakato, ʻi he tuʻunga maʻulalo ʻo e suka, pea toe fakalahi permeability, E. kololi B ngaahi selo ʻoku faʻa fakaʻaongaʻi ki hono ngaohi ʻo genetically engineered polotini.

Recombinant ivi

ʻOku mahuʻinga ʻaupito ʻa e Recombinant polotini (RProt) ʻi he ngaahi kolo manifold, kau ai ʻa e ngaohiʻanga kemikale, fakafaitoʻo, meʻafua, faitoʻo ʻo e tangata mo e monumanu, ngoue, meʻakai pea pehe ki he ngaahi ngaueʻanga ʻoku maumau.
ʻOku fie maʻu ki hono fokotuʻu ʻo recombinant ivi ke fakaʻaongaʻi ha founga ke fakahaaʻi ʻaki. ʻI hono fakahaaʻi ko ia ʻo e ngaahi founga toʻotoʻo ki hono ngaohi ʻo recombinant DNA, ʻe lava ke fakaʻaongaʻi fakatouʻosi ʻa e prokaryotic mo e ngaahi selo eukaryotic. ʻOku lahi taha hono fakaʻaongaʻi ʻo e hangatamaki toʻotoʻo ki hono fakahaaʻi ʻo e ivi koeʻuhi ko e ngaahi moʻoniʻi meʻa hange ko e totongi maʻamaʻa, faingofua scalability mo e ngaahi tuʻunga faingofua ʻo e mitia, mammalian, ʻio, algae, ʻinisekite mo e ngaahi founga taʻetotongi toʻotoʻo. Ko e faʻahinga ivi, ʻekitiviti ʻoku lele lelei, pea pehe ki he ngaahi meʻa ʻoku fie maʻu ke fakahaaʻi ʻaki ʻa e ivi takiekina ki hono fili ʻo e polokalama toʻotoʻo ʻoku fakaʻaongaʻi ki hono fakahaaʻi ʻo e ivi.
Koeʻuhi ke lava ʻo fakahaaʻi recombinant ivi, kuo pau ke transfected ha meʻa toʻotoʻo ʻaki ha DNA vector ʻoku ʻi ai ʻa e sipinga ʻo recombinant DNA. ʻOku toki fakahoko leva ʻa e ngaahi selo transfected mo e template. Ko e ola ʻo e founga telefoni toʻotoʻo, ko e hiki ʻa e ngaahi selo pea liliu ʻa e ivi ʻo e tokanga, ʻo maʻu ai ʻa e ivi kuo fakataumuʻa ki ai.
ʻI he taimi ʻoku maʻu ai ʻa e polotini ʻi he matrix Mopaila, kuo pau ke lysed ʻa e toʻotoʻo (motuhia mo maumauʻi) ke tukuange ʻa e polotini. ʻI ha sitepu fakamaʻa kimui ange, ʻoku mavahevahe mo fakamaʻa ʻa e ivi.
Ko e fuofua ivi recombinant ʻoku fakaʻaongaʻi ʻi hono faitoʻo ko recombinant ngaahi homouni tangata ʻi he 1982. ʻI he ʻaho ni, ʻoku laka hake ʻi he faʻahinga ʻe 170 ʻo e recombinant ʻoku faʻu ʻi he funga ʻo e mamani ki he ngaahi faitoʻo fakafaitoʻo. ʻOku angamaheni ʻaki hono fakaʻaongaʻi recombinant polotini ʻoku fakaʻaongaʻi ʻi he faitoʻo, hange ko ʻeni recombinant homouni, interferons, interleukins, ngaahi moʻoniʻi meʻa, foʻi ngungu necrosis, toto clotting ngaahi moʻoniʻi meʻa, thrombolytic e faitoʻo konatapu, mo e enzymes ki hono ngaohileleiʻi ʻo e ngaahi mahaki lalahi hange ko e suka, dwarfism, myocardial infarction, congestive ʻikai ke ola lelei e mafu, mamatea apoplexy, uoua lahi, neutropenia, thrombocytopenia, anemia, mahaki (FF. Phuc V. Phami, ʻi he Omics tekinolosia mo e piokalafi-ʻenisinia, 2018)


Hielscher Ultrasonics supplies high-performance ultrasonic homogenizers from lab to industrial size.

Ngaue ultrasonics maʻolunga! ʻOku ʻufiʻufi ʻe he koloa ʻa Hielscher ʻa e tuʻunga kakato mei he fale fakatotolo fakakemi ultrasonicator ʻi he ngaahi ʻiuniti ʻi ʻolunga ki he ngaahi polokalama ultrasonic lalahi.