WO1991006553A1 - Adn de transglutaminase de type i - Google Patents

Adn de transglutaminase de type i Download PDF

Info

Publication number
WO1991006553A1
WO1991006553A1 PCT/US1990/006075 US9006075W WO9106553A1 WO 1991006553 A1 WO1991006553 A1 WO 1991006553A1 US 9006075 W US9006075 W US 9006075W WO 9106553 A1 WO9106553 A1 WO 9106553A1
Authority
WO
WIPO (PCT)
Prior art keywords
transglutaminase
dna
type
probe
mrna
Prior art date
Application number
PCT/US1990/006075
Other languages
English (en)
Inventor
Anton M. Jetten
Esther E. Floyd
Original Assignee
The United States Of America, Represented By The Secretary, United States Department Of Commerce
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.)
Filing date
Publication date
Application filed by The United States Of America, Represented By The Secretary, United States Department Of Commerce filed Critical The United States Of America, Represented By The Secretary, United States Department Of Commerce
Publication of WO1991006553A1 publication Critical patent/WO1991006553A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/1096—Transferases (2.) transferring nitrogenous groups (2.6)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers

Definitions

  • This invention relates to a DNA segment encoding type I transglutaminase. Furthermore, the present inven- tion relates to a method for detection of squamous cell carcinomas.
  • Transglutaminases (EC 2.3.2.13) are a family of calcium-dependent enzymes which catalyze an acyl-transfer reaction between peptide-bound gluta ine residues and primary amines (Folk, Ann. Rev. Biochem. 49:517-531 (1980) and Lorand, et al, Molec. Cell. Biochem. 58:9-35 ((1984) resulting either in the covalent incorporation into proteins of small, primary amines or in the cross-linking of proteins by the formation of e(gamma-glutamyl)lysine trans-peptide bonds.
  • transglutaminases include blood coagulation factor Xllla, which is distributed both intracellularly, and the intra- cellular enzymes: type II ("tissue”) transglutaminase and type I ("epidermal”) transglutaminase (Folk, Ann. Rev. Biochem. 49:517-531 (1980).
  • squamous differentiation represents an abnormal pathway of cellular differentiation which can be observed under conditions of vitamin A deficiency, hyperplasia and squamous metaplasia.
  • Type I transglutaminase activity is a marker for squamous cell differentiation. Increase in type I trans ⁇ glutaminase activity correlates with other characteristics of squamous cell differentiation in epithelial cells in culture. This increase in type I transglutaminase activi ⁇ ty is inhibited by retinoic acid which is known to inhibit the expression of the squamous phenotype but not terminal cell division (Jetten, el al, J. Biol. Chem. 261: 15097 (1986)).
  • the present invention embodies the use of recombi- nant DNA methods to provide a DNA segment encoding type I transglutaminase, and the use of that DNA for detection and identification of squamous cell carcinomas, analyses of disease processes, the study of the regulation of squamous cell differentiation and identification of DNA segments encoding other members of the transglutaminase family.
  • the present invention relates to a DNA segment having a sequence which encodes mammalian type I (epider ⁇ mal) transglutaminase. More specifically, for example, the present invention relates to the DNA segment that encodes the rabbit type I transglutaminase which is clone pTG-7.
  • the DNA segment of the present invention was enabled by the invention of a DNA probe which is comple ⁇ mentary to mRNA sequences encoding the conserved sequence of a ino acids at the active site of known transglutamin- ase enzymes.
  • the DNA segment of the present invention has uses in bioassay for the expression of a gene encoding type I transglutaminase and in a method for the detection or identification of squamous cell carcinomas.
  • the invention also relates to a method for isola ⁇ tion of complementary DNA corresponding to a new member of the transglutaminase family of enzymes. All publications mentioned hereunder are hereby incorporated herein by reference.
  • Figure 1 shows in the upper sequence the conserved amino acid sequence found at the active sites of guinea pig liver transglutaminase type II and human blood coagu ⁇ lation factor Xllla.
  • the middle sequence shows all possible mRNA sequences encoding the conserved amino acid sequence above.
  • the lower sequence shows the 16-fold degenerate oligonucleotide probe for the transglutaminase type I gene.
  • Bases are as customary A for adenosine, T for thymidine, C for cytosine and G for guanosine.
  • I is for deoxyinosine which allows hybrid formation with any of the four usual bases on the complementary strand.
  • Figure 2 shows the restriction maps of cDNA clones pTG-7 and pTG-15 where R, T and X stand for EcoRI, Sstl and Xhol restriction sites respectively.
  • the solid bars indicate the restriction fragments to which the mixed oligonucleotide probe hybridized.
  • Figure 3 shows the partial nucleotide sequence of pTG-7.
  • Figure 4 compares the partial amino acid sequence of pTG-7 with the amino acid sequence of other trans ⁇ glutaminases. Asterisks indicate points of sequence homology among all three proteins and dots indicate points of homology between pTG-7 and one of the other two se ⁇ quences .
  • the present invention relates to a DNA probe having sequences which are complementary to all possible mRNA sequences encoding the conserved sequence of amino acids at the active site of two known transglutaminases, namely guinea pig liver transglutaminase type II and human blood factor Xllla.
  • the present invention is also related to a method for isolation of DNA segments encoding new members of the transglutaminase family.
  • the invention relates to a bioassay for the expression of a gene encoding type I transglutaminase for the identifica- tion or detection or squamous cell carcinomas.
  • a cDNA clone was derived from rabbit tracheal epithelia (RbTE) using a probe whose sequence was based on the conserved human factor Xllla and guinea pig liver type I transglutaminase sequences and was able to selectively identify type I transglutaminase message in human cells. From this it would be clear to one of ordinary skill in the art that the present clone could be used to isolate human or any other mammalian transglutaminase according to standard methods of this invention. One skilled in the art knows it is not necessary to use the entire cDNA clone as a probe.
  • the minimum probe for detecting transglutaminases would be a 12 nucleotide-long oligonucleotide comprising all sequen ⁇ ces encoding 4 consecutive amino acids.
  • An oligonucleo- tide of 13 to 15 nucleotides may be necessary in some cases for sufficient annealing to occur to detect comple ⁇ mentarity.
  • the original oligonucleotide probe is also useful for cloning mammalian transglutaminase genes since it contains the eight amino acid sequence conserved in known transglutaminase. This aspect is exemplified by the isolation of the cDNA encoding rabbit type I trans ⁇ glutaminase as described in the examples.
  • the present invention relates to antibodies to the type I transglutaminase. It is routine for one of ordinary skill in the art to generate antibod ⁇ ies against the type I transglutaminase by, for example, using a peptide corresponding to the sequence determined from the DNA segment that encodes mammalian type I trans- glutaminase of the present invention. Methods for this are well known in the art, such as those in Culler et al, Brain Research Bulletin, 17:219(1986). The antibodies are used in screening assays such as ELISA to identify samples expressing high type I transglutaminase activity.
  • the amino acid sequence of type I trans ⁇ glutaminase is deduced from the cDNA clone of the present invention and a corresponding synthetic peptide is pro ⁇ substituted by Bachem.
  • the immunogen is formed by conjugating the peptide to a carrier such as thioglobulin.
  • a carrier such as thioglobulin.
  • approximately 1000 ⁇ g of the peptide is emulsified with Freund's complete adjuvant.
  • About 250 ⁇ g of the peptide is injected into a rabbit, with a booster injection of 135 ⁇ g/rabbit occurring 10 days later.
  • About 20 days after the second injection the rabbit is bled from the marginal ear vein and the sera containing the type I transglutaminase antibodies is collected
  • a bioassay for determining levels of expression a gene encoding type I transglutaminase is used to detect or identify squamous cell carcinomas.
  • Squamous cell carcino ⁇ mas are detected and identified by a method comprising the steps of screening tissues containing potential carcinoma cells, such as lung tissue, with methods including i situ hybridization or Northern blotting, using the type I transglutaminase DNA of this invention as a probe.
  • the availability of the type I transglutaminase cDNA makes possible early detection of carcinomas.
  • squamous differentiation represents an abnormal pathway of cellular differentiation in tissues such as tracheobronchial epithelium
  • detection of high levels of type I trans ⁇ glutaminase mRNA is an early indicator of carcinoma conditions.
  • standard methods such as radioactive labeling of the probe or immunofluorescence are used.
  • novel DNA segments encoding for new members of trans- gluta inase family are identified and isolated by screen ⁇ ing a cDNA library made from poly(A+) RNA of a cell preferably expressing high quantities of the trans ⁇ glutaminase protein of interest with an oligonucleotide.
  • the oligonucleotide is complementary to all possible mRNA sequences encoding the conserved sequence of amino acids at the active sites of known transglutaminase enzymes, as exemplified by the probe sequence shown in Figure 1.
  • Clones to which the probe hybridizes can be further characterized by common procedures including restriction mapping, partial sequencing and cross-hybridization.
  • a method for the detection or identification of squamous cell carcinomas comprises the steps of using the bioassay of the present invention to identify the level of type I transglutaminase expression and comparing that level to a threshold level of type I transglutaminase expression.
  • the threshold level is determined by taking the average level of type I transglutaminase expression in about 5 standard epithelial cells. Tissues or cells with levels of type I transglutaminase expression above the threshold level are identified as squamous cell carcinomas.
  • One aspect of the present invention is a DNA segment having a sequence which encodes mammalian type I transglutaminase.
  • Another aspect of the present invention is a DNA probe having a sequence which is complementary to all mRNA sequences encoding the conserved sequence of amino acids at the active site of known transglutaminases. These aspects are illustrated here with the isolation of transglutaminase clones from rabbit cells using this DNA probe.
  • the 23-nucleotide, 16-fold degenerate mixed oligonucleotide probe used to screen the cDNA library was synthesized by OCS Laboratories (Denton, TX) . Sequencing primers were commercially available or were synthesized using a Beckman System 1 Plus DNA synthesizer.
  • This mixture of 16 oligonucleotides was used to screen a LambdaZap library of cDNAs derived from calcium- treated, squamous-differentiated RbTE cells.
  • RbTE cells were isolated and grown in primary cultures as has been previously described (Jetten el al, J. Biol. Chem. 261:15097 (1986).
  • Poly(A)+RNA isolated from squamous- differentiated RbTE cells grown in the presence of 2mM calcium, was utilized to prepare a cDNA library in the vector LambdaZap.
  • the library contained 4.8 x 10 6 primary plaques .
  • the transglutaminase mixed-oligonucleotide probe was labeled with 32 P using polynucleotide kinase, and prepared for use in hybridization reactions as described (Berent et al, Biotechniques 3:208-220 (1985)).
  • pgmTG700 was digested with HindiII and transcribed from the T7 promot ⁇ er.
  • pTG-7 was digested with Sail and tran ⁇ scribed from the T3 promoter.
  • pTG-7 was digested with EcoRI, and the 2.7 kb cDNA insert was isolated and labeled by nick-translation.
  • LambdaZap recombinants were plated at a density of 1 x 104 phage per 150 mm plate.
  • Replicate filters of each plate were prepared as in (Benton et al, Science 196:180- 182 (1977)) and hybridized to 32 P-labeled mixed oligo- nucleotides probe essentially as described in (Ausubel et al, 1987, Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York) with the final wash at 50 in 6X SSC plus 0.05% sodium pyrophosphate.
  • group 1 contained the clones pTG-4, pTG-7, pTG-9 and pTG-10 and group 2, which contained a single clone pTG-15.
  • Group 1 clones contained inserts ranging in size from approximately 1.7 kb to 2.6 kb. These four clones gave similar restriction maps and their cDNA inserts hybridized strongly to the pTG-7 insert. Sequencing of several hundred bases at the ends of each clone pinpointed the poly(A) tracts, establishing the 5' to 3' orientation of the clones. Clone pTG-7 was selected for further analysis. pTG-15 contained an approximately 3.5 kb insert which gave a restriction map that was very different from that of the pTG-7-like clones (see Figure 2).
  • pTG-15 did not cross-hybridize with the insert of pTG-7. Further, sequence analysis of pTG-15 at either end of the insert failed to reveal substantial homology with the pTG-7-like clones. Most likely pTG-15 represents the product of a transcriptional unit which is distinct from that of pTG-7.
  • One aspect of the present invention is a DNA segment, clone pTG-7 having a sequence which encodes a mammalian type I transglutaminase. This aspect is illus ⁇ trated below with the identification of clone pTG-7 encoding rabbit type I transglutaminase.
  • Clones were analyzed by Southern blotting with hybridization of the blots to the mixed oligonucleotide probe, to delineate the region of each clone to which the mixed probe hybridized.
  • DNA blots were hybridized to nick-translated probes (2-4 x 10 8 cpm/ ⁇ g) essentially as described elsewhere (Smits et al, Molec. Cell. Biol. 7:4017 (1987)).
  • DNA blots were hybridized to the 32 P-labeled mixed oligonucleotide probe essentially as described for plaque hybridizations.
  • pTG-7 revealed a region, on the non- coding strand, which exhibited perfect homology to one of the 16 oligonucleotide in the mixed probe, namely 5'- GCAAAIACCCAGACTGICCATA-3' .
  • the complementary region on the coding strand of pTG-7 lies within an open reading frame and encodes the conserved, eight amino acid sequence found at the active site of both human factor Xllla and guinea pig liver transglutaminase type II . Further comparison of the translated amino acid sequence of the 1.1 kb Xhol fragment of pTG-7 with the sequences of human factor Xllla showed regions of substantial homology at the amino acid level as seen in Figure 4.
  • HNEK cells normal human epidermal keratinocytes
  • mRNA levels in normal human epidermal keratinocytes which express moderate levels of both type I and type II transglutaminase proteins
  • NHEK cells were obtained from Clonetics Corp. (Palo Alto, CA) and grown in KGM medium as previously described (Jetten et al, J. Invest. Dermatol. 92:203 (1989)) .
  • northern blots were prehybridized for 4-12 hours at 55°C in a buffer containing 50% for- mamide, 5X SSPE, 5X Denhardt's, 1% SDS and 250 ⁇ g/ml sheared salmon sperm DNA. Hybridization was overnight at 55°C in prehybridization buffer containing 500 ⁇ g/ml yeast tRNA plus the probe (approximately 50 ng/ml). The blots were washed at a final stringency of 55°C in 0.IX SSC, 0.1% SDS.
  • RNA blots were hybridized to nick-translated probes at 42°C as described for DNA blots, and washed at a final stringency of 60°C in 2X SSC, 0.5% SDS.
  • Hybridization signals detected by autoradiography were quantitated by scanning with a BIORAD model 620 video densitometer and the scans analyzed with the 1-D Analyst software package.
  • Hybridization of 32 P-pTG-7 to a northern blot of poly(A)+ RNA (2 ⁇ g/lane) isolated from cultured prolifera ⁇ tion (P) or calcium-treated squamous-differentiated (C) NHEK cells revealed a single size of mRNA migrating at approximately 3.6 kb.
  • pgmTG700 which encodes a 691 base pair portion of the cDNA for mouse acrophage transglutaminase type II (Chiocca, et al, J. Biol. Chem. 263:11584 (1988) a single size of mRNA migrat- ing at approximately 4.4 kb was identified.
  • Clone pgmTG700 comprises the vector pGEM3Z (Promega). This insert in the vector pBR328 has been previously described in (Chiocca, et al, J. Biol. Chem. 263:11584 (1988).
  • the 3.6 to 4.7 kb mRNAs also exhibited differing patterns of expression in NHEK cells under the various culture conditions tested. These results indicate that the clone pTG-7 encodes type I, or epidermal, trans ⁇ glutaminase.
  • Example 3 Regulation of the mRNA for Transglutaminase Type I .
  • Expression of the squamous phenotype by RbTE cells is accompanied by a dramatic increase in transglutaminase type I activity; this increase is further enhanced when cells are cultured in the presence of calcium (Jetten et al, J. Biol. Chem. 261:15097 (1986)).
  • Retinoid which inhibit expression of the squamous phenotype but not terminal cell division (Jetten Environ. Health Perpsect. 80:149-160 (1989)), block this increase in type I trans ⁇ glutaminase.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biomedical Technology (AREA)
  • Hospice & Palliative Care (AREA)
  • Oncology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un segment ADN qui code la transglutaminase de type I mammifère. Elle concerne également des procédés permettant d'isoler de nouveaux membres de la famille transglutaminase et de détecter ou identifier des carcinomes de cellules squameuses.
PCT/US1990/006075 1989-10-24 1990-10-24 Adn de transglutaminase de type i WO1991006553A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US42588789A 1989-10-24 1989-10-24
US425,887 1989-10-24

Publications (1)

Publication Number Publication Date
WO1991006553A1 true WO1991006553A1 (fr) 1991-05-16

Family

ID=23688451

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1990/006075 WO1991006553A1 (fr) 1989-10-24 1990-10-24 Adn de transglutaminase de type i

Country Status (2)

Country Link
AU (1) AU6608590A (fr)
WO (1) WO1991006553A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993013207A2 (fr) * 1991-12-31 1993-07-08 Zymogenetics, Inc. Nouvelles transglutaminases humaines
WO1994010296A1 (fr) * 1992-11-03 1994-05-11 Oklahoma Medical Research Foundation Gene de la transglutaminase
US5958752A (en) * 1993-04-30 1999-09-28 The United States Of America As Represented By The Department Of Health And Human Services Nucleic acid molecules encoding human trichohyalin and use thereof

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
AGRICULTURAL BIOLOGICAL CHEMISTRY, Vol. 51, No. 3, issued 1987, IKURA et al., "Cloning of cDNA Coding for Guinea Pig Liver Transglutaminase", pages 957-961. *
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, Vol. 83, issued November 1986, GRUNDMANN et al., "Characterization of cDNA Coding for Human Factor XIIIa", pages 8024-8028. *
T. MANIATIS et al., "Molecular Cloning, A Laboratory Manual", published 1982 by MACGRAW-HILL (N.Y.), see pages 224-234, 320-321, and 330-333. *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993013207A2 (fr) * 1991-12-31 1993-07-08 Zymogenetics, Inc. Nouvelles transglutaminases humaines
WO1993013207A3 (fr) * 1991-12-31 1993-09-30 Zymogenetics Inc Nouvelles transglutaminases humaines
US5514579A (en) * 1991-12-31 1996-05-07 Zymogenetics, Inc. Human transglutaminases
US5952011A (en) * 1991-12-31 1999-09-14 Zymogenetics, Inc. Human transglutaminases
WO1994010296A1 (fr) * 1992-11-03 1994-05-11 Oklahoma Medical Research Foundation Gene de la transglutaminase
US5726051A (en) * 1992-11-03 1998-03-10 Oklahoma Medical Research Foundation Transglutaminase gene
US5958752A (en) * 1993-04-30 1999-09-28 The United States Of America As Represented By The Department Of Health And Human Services Nucleic acid molecules encoding human trichohyalin and use thereof

Also Published As

Publication number Publication date
AU6608590A (en) 1991-05-31

Similar Documents

Publication Publication Date Title
Amara et al. Alternative RNA processing in calcitonin gene expression generates mRNAs encoding different polypeptide products
Lin et al. Molecular cloning and characterization of plastin, a human leukocyte protein expressed in transformed human fibroblasts
Lawton et al. Molecular cloning of plant transcripts encoding protein kinase homologs.
Floyd et al. Regulation of type I (epidermal) transglutaminase mRNA levels during squamous differentiation: down regulation by retinoids
Heiss et al. X-linked dyskeratosis congenita is caused by mutations in a highly conserved gene with putative nucleolar functions
Gisselbrecht et al. Frequent c-fms activation by proviral insertion in mouse myeloblastic leukaemias
Pringle et al. In situ hybridization of immunoglobulin light chain mRNA in paraffin sections using biotinylated or hapten‐labelled oligonucleotide probes
Linton et al. Dual bidirectional promoters at the mouse dhfr locus: cloning and characterization of two mRNA classes of the divergently transcribed Rep-1 gene
Swinkels et al. Characterization of the gene for the microbody (glycosomal) triosephosphate isomerase of Trypanosoma brucei.
US5635354A (en) Method for describing the repertoires of antibodies (Ab) and of T-cell receptors (TcR) of an individual's immune system
Ramamoorthy et al. Proton/peptide cotransporter (PEPT 2) from human kidney: functional characterization and chromosomal localization
Mahon et al. Structure and expression of the egg-laying hormone gene family in Aplysia
de Veer et al. IFI60/ISG60/IFIT4, a new member of the human IFI54/IFIT2 family of interferon-stimulated genes
Sunnerhagen et al. The effect of aspartate hydroxylation on calcium binding to epidermal growth factor-like modules in coagulation factors IX and X.
US5618678A (en) Methods for detecting PDGF agonist or antagonist activity using PDGF .alpha.
US5624801A (en) Methods of identifying human osteoclast-specific and related genes
Zomerdijk et al. Antigenic variation in Trypanosoma brucei: a telomeric expression site for variant-specific surface glycoprotein genes with novel features
Abrams et al. Molecular cloning of the microfibrillar protein MFAP3 and assignment of the gene to human chromosome 5q32–q33. 2
Ruthertord et al. The murine 2-5A synthetase locus: three distinct transcripts from two linked genes
Stengel et al. Different chromosomal localization of two adenylyl cyclase genes expressed in human brain
Chaudhuri et al. The gene for a novel protein, a member of the protein disulphide isomerase/form I phosphoinositide-specific phospholipase C family, is amplified in hydroxyurea-resistant cells
WO1991006553A1 (fr) Adn de transglutaminase de type i
Butler et al. Mapping of the mouse serum amyloid A gene cluster by long-range polymerase chain reaction
Amler et al. Identification and characterization of novel genes located at the t (1; 15)(p36. 2; q24) translocation breakpoint in the neuroblastoma cell line NGP
Planck et al. Modulation of hnRNP A1 protein gene expression by epidermal growth factor in Rat-1 cells

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IT LU NL SE

NENP Non-entry into the national phase

Ref country code: CA