WO1990002188A1 - SUBSTITUTION PHAGE VECTOR μSK15 FOR CONSTRUCTION OF kDNA STRUCTURE GENE LIBRARIES - Google Patents

SUBSTITUTION PHAGE VECTOR μSK15 FOR CONSTRUCTION OF kDNA STRUCTURE GENE LIBRARIES Download PDF

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Publication number
WO1990002188A1
WO1990002188A1 PCT/SU1989/000230 SU8900230W WO9002188A1 WO 1990002188 A1 WO1990002188 A1 WO 1990002188A1 SU 8900230 W SU8900230 W SU 8900230W WO 9002188 A1 WO9002188 A1 WO 9002188A1
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WO
WIPO (PCT)
Prior art keywords
thousand
phage
size
nuclei
fragment
Prior art date
Application number
PCT/SU1989/000230
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English (en)
French (fr)
Russian (ru)
Inventor
Evgeny Reanadovich Zabarovsky
Elena Gennadievna Zagorskaya
Lev Lvovich Kiselev
Original Assignee
Institut Molekulyarnoi Biologii Imeni V.A.Engelga Rdta Akademii Nauk Sssr
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
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Application filed by Institut Molekulyarnoi Biologii Imeni V.A.Engelga Rdta Akademii Nauk Sssr filed Critical Institut Molekulyarnoi Biologii Imeni V.A.Engelga Rdta Akademii Nauk Sssr
Priority to DE19893990972 priority Critical patent/DE3990972T1/de
Publication of WO1990002188A1 publication Critical patent/WO1990002188A1/ru
Priority to GB9009525A priority patent/GB2230012A/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C12N15/73—Expression systems using phage (lambda) regulatory sequences
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA

Definitions

  • the project provides the ability to consume libraries of CDD genes using a large number of tests
  • ⁇ 392 ⁇ ', ⁇ z ⁇ ⁇ 514 (r ⁇ , t ⁇ ) zi ⁇ 44, zi ⁇ ⁇ 58, 30 ⁇ as ⁇ ⁇ ( ⁇ as I ⁇ ) 6 ⁇ a1 ⁇ 2, e a1 ⁇ 22, those "L ⁇ , 1; g ⁇ 55 0 . ⁇ .
  • the claimed vector allows you to receive libraries of industrial genes for the use of industrial and electronic recordings, which is a significant gain of interest in the study. This is the only
  • the claimed vector allows, using biochemical and genetic selection, effective
  • the inventive vector allows the possibility to carry out the expressions of CDD and the translation of M in the cells ⁇ . a mixture that carries a mutation of zi ⁇ C and / or zi ⁇ &, zi ⁇ D, zi ⁇ ⁇ . (" ⁇ réelle"). With this option, you can save libraries as well as traditional methods, as well as modifiable methods
  • the claimed vector makes it easy to amplify the propagating gene using the Dreambook.
  • the inventive part I-3' ⁇ 5 allows you to easily get rid of the “shoulders” of the phage L and transfer the insert to the plasmid form, and if the carcass is infected.
  • SOSHE soderu- protects the plate
  • the phobom-assistant of the UZ is located in the proteins of the phage OZ and is transferred to the single unit
  • FIG. 2 illustrates the reduction of the 5–8 band
  • Fig. 3 depicts the direct compression of the vector ⁇ ⁇ from phasmid ⁇ 5 ⁇
  • Fig. 4 depicts the end of the phasing of the ages 15 at 5-5
  • Fig. 5 depicts the scheme of copying the library of genes KDV in L 5–5 using the use of KAT ⁇
  • Fig. 6 depicts the general design of a CD library in LP5 using the partial access of 20 20 kix cards
  • Fig. 7 depicts the design of the KD ⁇ library in _D 5 comparativelyatter5 with the use of special links.
  • BEST MODE FOR CARRYING OUT THE INVENTION The claimed phrased version L 5 ⁇ 5 is constituted on the 25th of the following scheme. Initially, they carry out the formation of the vector 5–8 (Figs. A, b and 2).
  • I remove and clean the elec- tric element in the light-melting unit with the size of 1.86 thousand.
  • the C79 promotes the production of waste and removes the constituent that contains the co-participation.
  • ⁇ 5 ⁇ 2 is hydrolyzed and in conjunction with the group containing -
  • Partition 5–6 hydrates the distant component, which allows for the loss of other components in the process
  • 35 shtamgl Z. 109 which are sown on a medium containing • ampicillin, X- ⁇ , ⁇ .
  • a medium containing • ampicillin, X- ⁇ , ⁇ Of the small species, in particular, with amcycillin, cells and excretory cells are dysfunctional.
  • ⁇ For industrial purposes, ⁇ certify ⁇ , ⁇ - ⁇ , ⁇ ⁇ , ⁇ Vietnamese I, - 8 - in ⁇ ⁇ , ⁇ réelle ⁇ and analyze ⁇ . From the analysis of the identified data, a partition of 5–8 is distinguished, which is a quick hydrostatic access to the simplest section of the ISS and allows for the participation of the Russian Federation in the event of a permanent infection.
  • the phasmids secrete the lobe from ⁇ 2 ⁇ .
  • the loss of zone 2 is due to the loss of distance of 2 without participation in the recognition of social and social problems outside of the public domain. Then, the reduction of the recognition site of the disease is recognized in section 5-2.
  • the phasing factor is 5 ⁇ 2 ⁇
  • the league is introduced into the radial cage. sostokh d. 109, and disperse on agar with ampsitshinom. With a large size of prints on nitrous cellulose filters and converts hybridsing with the machine, it is possible to receive a good result. Battery from the cell
  • I receive a flatbed ⁇ 4 ⁇ - ⁇ .
  • the ⁇ 4 ⁇ plasma torch is used to stabilize the test and remove the reactant by heating 15 cables at 65 ° ⁇ . Then D phage I also expand and ⁇ agment
  • the 10th case (Fig. 4), called ⁇ -- ⁇ , is used in the next stage, at the completion of the processing of ⁇ 5 ⁇ 5.
  • ⁇ -- ⁇ For the plasti ⁇ 4 ⁇ - ⁇ , they hydrolyze the products of Soson and ⁇ a ⁇ and the reaction restores by heating 15 cables at 65 ° ⁇ . D ⁇ promotes cultivation
  • the claimed phrasal vector L 5 ⁇ 5 may be used for the purpose of obtaining libraries for the permanent method with the use of the site - the site.
  • Such and its recombinants may be converted to a club and other places.
  • the integral axial separation of the phage on the mains plugs, the resis- tant to replicate the phage L (for example, ⁇ , ⁇ . ⁇ . ⁇ ) is inactive for 5-10 am-
  • the percentage of recombinants in the library, obtained in L 5–5, is easy to target at random by re-injecting plaques on a gaz ⁇ 359 or using a colored gas
  • the proposed dual lawn technology allows for the delivery of these disadvantages.
  • the analyzed phages are incubated with the cells of the ⁇ .social strain ⁇ 392 and, after the recovery of the 0.6 ⁇ -th agaris, is sown in the cups of the Petri.
  • this lawn is sown on lawn; the cage 0359 also - 16 - in the 0.6 ⁇ -th b-agar.
  • recombinant phages of the dagota produce useful plaques, while non-recombinant phages are muddy, which is easily visualized.
  • the claimed phrase vector is an unqualified replacement vector, which can be used for the purpose of downloading the library.
  • the claimed vector implies:
  • s ⁇ de ⁇ z haschem 50 m ⁇ ⁇ is- ⁇ S ⁇ ( ⁇ 7,5), 10 m ⁇ m ⁇ 2, 10 m ⁇ tsi ⁇ i ⁇ ei ⁇ a 3 m ⁇ atsen ⁇ zin ⁇ i ⁇ s ⁇ a ⁇ a 25 etsinits a ⁇ ivn ⁇ s ⁇ i D ⁇ ligase ⁇ aga ⁇ 4 m ⁇ g and 50 / ml summa ⁇ n ⁇ y D ⁇ .
  • the league was won into
  • the design which is conveniently flashed, is a quick-turnable quick-disconnect switch (fig. ⁇ ), and the above-mentioned ciphery wave is used for the most complete. With the aim of decreasing the size of 5, the burner will hydrate the rest of the product and at the same time (for 5 activity units), and
  • the event is an ongoing event, an electrical process from a fusible plant after an electric power plant.
  • 2 xi D ⁇ 5 ⁇ 2 ⁇ a ⁇ zhe gits ⁇ lizuyu ⁇ v ⁇ P and e ⁇ ⁇ e ⁇ a ⁇ a ⁇ ligi ⁇ uyu ⁇ with ⁇ agmen ⁇ m
  • s ⁇ tse ⁇ z haschim s ⁇ z-uchas ⁇ in bu ⁇ e ⁇ e
  • s ⁇ tse ⁇ z haschem 50 m ⁇ ⁇ is- ⁇ S ⁇ ( ⁇ 7,5), 10 m ⁇ ⁇ s ⁇ 2, Yu m ⁇ tsi ⁇ i ⁇ ei ⁇ a 3 m ⁇ atsen ⁇ zin ⁇ i ⁇ s ⁇ a ⁇ a ,
  • na ⁇ dyascheg ⁇ sya is ⁇ lilin- ⁇ e ⁇ a, ⁇ agmen ⁇ m ⁇ len ⁇ va in ⁇ isu ⁇ s ⁇ vii ⁇ and ⁇ .E ⁇ ⁇ e ⁇ a ⁇ a ⁇ ⁇ b ⁇ aba ⁇ yvago ⁇ D ⁇ ligaz ⁇ y ⁇ aga ⁇ 4 (5 units a ⁇ iv- n ⁇ s ⁇ i) in 50 m ⁇ l ⁇ as ⁇ v ⁇ a, s ⁇ de ⁇ zhascheg ⁇ 50 m ⁇ ⁇ is- ⁇
  • the phase-out factor combines the plague component ⁇ RB322 with the onset of the onset of replication and the gene for susceptibility to arterial hypertrophy, due to the
  • 25 bacteria are produced on the chaffs of Petri with 2 $ agar, containing 25 g / ml agglicycillin, 0.1 G ⁇ and 0.1 $ X-a ⁇ .
  • ⁇ - ⁇ This section, called ⁇ - ⁇ , is used for processing phage-vectors _ /? ⁇ 7 (fig. ⁇ ).
  • 0.5 mcg for use in patients with disruption of ⁇ and the enzyme inactivate by ingestion.
  • 3 mcg ⁇ ⁇ ⁇ 5 ⁇ 9 hydrolyzes the process of gas and food processing and inactivates the components by heating and purifying them from 20 relays-free for 600
  • the mixture is mixed (weight ratio of two vectors and a genus of 2: 1) and the ligament is mixed with a total concentration of 250 mg / ml.
  • 25 I • 10 mcg of the league DU are installed in the proteins of the phage ⁇ ⁇ - ⁇ réelle and 1/500 of the obtained phage particles are spread on the lawn Albany.
  • p. 359 is just not out of the ordinary, so in general, recombinant phages are over 95. From 6 and 30 randomly collected plaques from the lawn ⁇ 392 in 10 ml of b - mediums from ⁇ 392 phages were grown and removed from them.

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Virology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
PCT/SU1989/000230 1988-08-31 1989-08-29 SUBSTITUTION PHAGE VECTOR μSK15 FOR CONSTRUCTION OF kDNA STRUCTURE GENE LIBRARIES WO1990002188A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
DE19893990972 DE3990972T1 (de) 1989-08-29 1989-08-29 Phagen-vektor der substitution (lambda)sk15 zum konstruiereen von bibliotheken der sturkturgene kdns
GB9009525A GB2230012A (en) 1988-08-31 1990-04-27 Substitution phage vector lambda sk15 for construction of kdna gene libraries

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
SU884478259A SU1650699A1 (ru) 1988-08-31 1988-08-31 Фаговый вектор замещени @ К15 дл конструировани библиотек структурных генов к ДНК
SU4478259/31 1988-08-31

Publications (1)

Publication Number Publication Date
WO1990002188A1 true WO1990002188A1 (en) 1990-03-08

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ID=21397371

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/SU1989/000230 WO1990002188A1 (en) 1988-08-31 1989-08-29 SUBSTITUTION PHAGE VECTOR μSK15 FOR CONSTRUCTION OF kDNA STRUCTURE GENE LIBRARIES

Country Status (6)

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JP (1) JPH03505402A (enrdf_load_stackoverflow)
DE (1) DE3990972C2 (enrdf_load_stackoverflow)
GB (1) GB2230012A (enrdf_load_stackoverflow)
SE (1) SE465577B (enrdf_load_stackoverflow)
SU (1) SU1650699A1 (enrdf_load_stackoverflow)
WO (1) WO1990002188A1 (enrdf_load_stackoverflow)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU694269B2 (en) * 1994-09-22 1998-07-16 Merck Sharp & Dohme Corp. Dna encoding human papillomavirus type 6a

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
"DNA Cloning", Vol. II, Pod Redaktsiei D.M. GLOVER, 1985, IRL PRESS, (Oxford), pages 49-78. *
BIOCHEMISTRY, Vol. 26, No. 6, 1987, (American Chemical Society), JANG H. HAN et al., "Isolation of Full-Length Putative Rat Lysophospholipase cDNA Using Improved Methods for mRNA Isolation and cDNA Cloning", pages 1617-1624. *
NUCLEIC ACIDS RESARCH, Vol. 15, No. 15, 1987, (IRL Press Limited, Oxford, GB), JANG H. HAN et al., "Lambdagt22, an Improvedlambda Vector for the Directional Cloning of Full-Length cDNA", page 6304. *

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Publication number Publication date
GB2230012A (en) 1990-10-10
JPH03505402A (ja) 1991-11-28
GB9009525D0 (en) 1990-07-25
DE3990972C2 (enrdf_load_stackoverflow) 1992-12-24
SE9001560D0 (sv) 1990-04-30
SE9001560L (sv) 1990-04-30
SE465577B (sv) 1991-09-30
SU1650699A1 (ru) 1991-05-23

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