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Secreted and transmembrane polypeptides and nucleic acids encoding the same

a technology of transmembrane polypeptides and nucleic acids, applied in the field of identification and isolation of novel dna, can solve the problems of not manifesting clinical symptoms, problems with these diagnostic techniques, and limited mcp-1 targets to monocytes and basophils

Inactive Publication Date: 2003-09-11
GENENTECH INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the targets of MCP-1 are limited to monocytes and basophils.
Problems exist with these diagnostic techniques.
First, patients may not manifest clinical symptoms at early stages of disease.
Second, serological tests do not always differentiate between invasive diseases and genetic syndromes.
However, the majority of mammalian Ig-CAMs appear to be too widely expressed to specify navigational pathways or synaptic targets suggesting that other CAMs, yet to be identified, have role in these more selective interactions of neurons.

Method used

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  • Secreted and transmembrane polypeptides and nucleic acids encoding the same
  • Secreted and transmembrane polypeptides and nucleic acids encoding the same
  • Secreted and transmembrane polypeptides and nucleic acids encoding the same

Examples

Experimental program
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example 1

[0506] Extracellular Domain Homology Screening to Identify Novel Polypeptides and cDNA Encoding Therefor

[0507] The extracellular domain (ECD) sequences (including the secretion signal sequence, if any) from about 950 known secreted proteins from the Swiss-Prot public database were used to search EST databases. The EST databases included public databases (e.g., Dayhoff, GenBank), and proprietary databases (e.g. LIFESEQ.TM., Incyte Pharmaceuticals, Palo Alto, Calif.). The search was performed using the computer program BLAST or BLAST-2 (Altschul et al., Methods in Enzymology 266:460-480 (1996)) as a comparison of the ECD protein sequences to a 6 frame translation of the EST sequences. Those comparisons with a BLAST score of 70 (or in some cases 90) or greater that did not encode known proteins were clustered and assembled into consensus DNA sequences with the program "phrap" (Phil Green, University of Washington, Seattle, Wash.).

[0508] Using this extracellular domain homology screen, ...

example 2

[0511] Isolation of cDNA clones by Amylase Screening

[0512] 1. Preparation of Oligo dT Primed cDNA Library

[0513] mRNA was isolated from a human tissue of interest using reagents and protocols from Invitrogen, San Diego, Calif. (Fast Track 2). This RNA was used to generate an oligo dT primed cDNA library in the vector pRK5D using reagents and protocols from Life Technologies, Gaithersburg, Md. (Super Script Plasmid System). In this procedure, the double stranded cDNA was sized to greater than 1000 bp and the SalI / NotI Tinkered cDNA was cloned into XhoI / NotI cleaved vector. pRK5D is a cloning vector that has an sp6 transcription initiation site followed by an SfiI restriction enzyme site preceding the XhoI / NotI cDNA cloning sites.

[0514] 2. Preparation of Random Primed cDNA Library

[0515] A secondary cDNA library was generated in order to preferentially represent the 5' ends of the primary cDNA clones. Sp6 RNA was generated from the primary library (described above), and this RNA was use...

example 3

[0535] Isolation of cDNA Clones Using Signal Algorithm Analysis

[0536] Various polypeptide-encoding nucleic acid sequences were identified by applying a proprietary signal sequence finding algorithm developed by Genentech, Inc. (South San Francisco, Calif.) upon ESTs as well as clustered and assembled EST fragments from public (e.g., GenBank) and / or private (LIFESEQ.RTM., Incyte Pharmaceuticals, Inc., Palo Alto, Calif.) databases. The signal sequence algorithm computes a secretion signal score based on the character of the DNA nucleotides surrounding the first and optionally the second methionine codon(s) (ATG) at the 5'-end of the sequence or sequence fragment under consideration. The nucleotides following the first ATG must code for at least 35 unambiguous amino acids without any stop codons. If the first ATG has the required amino acids, the second is not examined. If neither meets the requirement, the candidate sequence is not scored. In order to determine whether the EST sequenc...

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Abstract

The present invention is directed to novel polypeptides and to nucleic acid molecules encoding those polypeptides. Also provided herein are vectors and host cells comprising those nucleic acid sequences, chimeric polypeptide molecules comprising the polypeptides of the present invention fused to heterologous polypeptide sequences, antibodies which bind to the polypeptides of the present invention and to methods for producing the polypeptides of the present invention.

Description

FIELD OF THE INVENTION[0001] The present invention relates generally to the identification and isolation of novel DNA and to the recombinant production of novel polypeptides.BACKGROUND OF THE INVENTION[0002] Extracellular proteins play important roles in, among other things, the formation, differentiation and maintenance of multicellular organisms. The fate of many individual cells, e.g., proliferation, migration, differentiation, or interaction with other cells, is typically governed by information received from other cells and / or the immediate environment. This information is often transmitted by secreted polypeptides (for instance, mitogenic factors, survival factors, cytotoxic factors, differentiation factors, neuropeptides, and hormones) which are, in turn, received and interpreted by diverse cell receptors or membrane-bound proteins. These secreted polypeptides or signaling molecules normally pass through the cellular secretory pathway to reach their site of action in the extr...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/17C07H21/04C07K14/435C07K16/40C12N5/06C12N9/00C12P21/02C12P21/06G01N33/53
CPCA61K38/17C07H21/04C07K16/40C07K14/435G01N33/53C12P21/06C12N5/06C12N9/00C12P21/02Y02A50/30C07K14/47C07K14/705
Inventor ASHKENAZI, AVI J.BAKER, KEVIN P.BOTSTEIN, DAVID A.DESNOYERS, LUCEATON, DAN L.FERRARA, NAPOLEONEFONG, SHERMANGAO, WEI-QIANGGERBER, HANSPETERGERRITSEN, MARY E.GODDARD, AUDREYGODOWSKI, PAUL J.GURNEY, AUSTIN L.KLJAVIN, IVAR J.MATHER, JENNIE P.NAPIER, MARY A.PAN, JAMESPAONI, NICHOLAS F.ROY, MARGARET ANNSTEWART, TIMOTHY A.TUMAS, DANIELWATANABE, COLIN K.WILLIAMS, P. MICKEYWOOD, WILLIAM I.ZHANG, ZEMIN
Owner GENENTECH INC