Secreted and transmembrane polypeptides and nucleic acids encoding the same
a technology of applied in the field of secreted and transmembrane polypeptides and nucleic acids encoding the same, can solve the problems of loss of biological activity and possible changes in immunogenicity
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example 2
[0425] Isolation of cDNA Clones by Amylase Screening
[0426] 7. Preparation of Oligo dT Primed cDNA Library
[0427] 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 Sall / NotI linkered cDNA was cloned into XhoI / NotI cleaved vector. pRK5D is a cloning vector that has an sp6 transcription initiation site followed by an Sfil restriction enzyme site preceding the XhoI / NotI cDNA cloning sites.
[0428] 2. Preparation of Random Primed cDNA Library
[0429] 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
[0449] Example 3
[0450] Isolation of cDNA Clones Using Signal Algorithm Analysis
[0451] 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...
example 4
[0452] Isolation of cDNA Clones Encoding Human PRO Polypeptides
[0453] Using the techniques described in Examples 1 to 3 above, numerous full-length cDNA clones were identified as encoding PRO polypeptides as disclosed herein. These cDNAs were then deposited under the terms of the Budapest Treaty with the American Type Culture Collection, 10801 University Blvd., Manassas, Va. 20110-2209, USA (ATCC) as shown in Tabl 7 below.
8 TABLE 7 Material ATCC Dep. No. Deposit Date DNA26843-1389 203099 Aug. 4,1998 DNA30867-1335 209807 Apr. 28, 1998 DNA34431-1177 209399 Oct. 17, 1997 DNA38268-1188 209421 Oct. 28, 1997 DNA40621-1440 209922 Jun. 2, 1998 DNA40625-1189 209788 Apr. 21, 1998 DNA45409-2511 203579 Jan. 12,1999 DNA45495-1550 203156 Aug. 25, 1998 DNA49820-1427 209932 Jun. 2, 1998 DNA56406-1704 203478 Nov. 17, 1998 DNA56410-1414 209923 Jun. 2, 1998 DNA56436-1448 209902 May 27, 1998 DNA56855-1447 203004 Jun. 23, 1998 DNA56860-1510 209952 Jun. 9, 1998 DNA56862-1343 203174 Sep. 1, 1998 DNA56868-...
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