Identification of Molecular Sequence Signatures and Methods Involving the Same

Abstract

Novel means and methods for analyzing hybridization data derived from hybridization assays between a target nucleic acid and differently sequenced polynucleotide probes involve selecting probe sets that define reference sequences for sequence signatures and deriving useful data about the nature of the target nucleic acid molecule based on its hybridization to the probes. The methods are useful for determining whether the target contains a nucleic acid or polypeptide sequence signature, whether the target encodes a member of a gene family, or whether the target is derived from one of any number of genes.

1 . A method for determining whether a target molecule has a sequence from a gene family member comprising:
 providing a polynucleotide array comprising, for each of at least two different gene family members, a set of polynucleotide probes that define a reference nucleotide sequence from the gene family member;   generating hybridization data by performing a hybridization reaction between the target nucleic acid molecule and the probes in the sets and detecting hybridization between the target nucleic acid molecule and each of the probes in the sets; and   processing the hybridization data to determine whether the target nucleic acid has the reference sequence from one of the gene family members.   
     
     
         2 . The method of  claim 1  further comprising the step of selecting the target nucleic acid molecule by determining whether the target hybridizes to a nucleic acid probe that hybridizes to a gene encoding the gene family members. 
     
     
         3 . The method of  claim 1  wherein the step of processing is performed by a programmable digital computer. 
     
     
         4 . The method of  claim 1  wherein the polynucleotide array further comprises, for each of the gene family members, a probe set defining a highly conserved region of the gene and a probe set defining a highly variable region of the gene. 
     
     
         5 . The method of  claim 1  wherein the polynucleotide array further comprises, for each of the gene family members, probe sets defining at least two highly conserved regions of the gene and probe sets defining at least two highly variable regions of the gene. 
     
     
         6 . The method of  claim 1  wherein the reference nucleotide sequence codes for an amino acid sequence and the array further comprises probe sets defining the degenerate set of nucleotide sequences encoding the amino acid sequence. 
     
     
         7 . The method of  claim 1  further comprising the step of determining the nucleotide sequence of the target nucleic acid molecule if the target does not have the sequence of the region of a gene family member. 
     
     
         8 . A method of determining whether a nucleic acid in a sample is a member of a gene family, comprising:
 selecting a hierarchy of assay techniques comprising at least a first and second assay, said first assay being selected to provide a determination of a presence, absence, or variant of a first sequence signature and said second assay being selected to provide a determination of a presence, absence or variant of a second sequence signature; wherein at least one of said assays employs a high-density nucleic acid array;   analyzing said nucleic acid sample using said first assay; and   determining whether said nucleic acid is a member of said gene family based on the results of said first and second assays.   
     
     
         9 . The method of  claim 8 , wherein said first sequence signature is a highly conserved region of a gene family. 
     
     
         10 . The method of  claim 8 , wherein said second sequence signature is a non-conserved region of a gene family. 
     
     
         11 . The method of  claim 8 , further comprising determining the full length sequence of said nucleic acid sample. 
     
     
         12 . The method of  claim 8 , wherein said gene family is the TGF-beta family. 
     
     
         13 . The method of  claim 8 , wherein said first or second sequence signature is between 9 and 100 nucleotides in length. 
     
     
         13 . The method of  claim 13 , wherein said first or second sequence signature is between 18 and 30 nucleotides in length. 
     
     
         14 . A method of selecting clones for analysis comprising:
 providing a support having a variety of clones associated therewith;   exposing said support to one or more polynucleotides under low, medium, or high stringency conditions to permit at least some hybridization between said clones and said polynucleotides;   identifying said clones that hybridize with said polynucleotides; and   selecting at least one of said clones not identified in said identifying step for analysis.   
     
     
         15 . The method of  claim 14 , wherein said support is a high-density nucleic acid array. 
     
     
         16 . A method of narrowing a sample for analysis, comprising,
 providing a sample containing nucleic acids;   analyzing whether said sample contains a sequence signature using a high-density nucleic acid array; and   further analyzing said nucleic acid sample only if said sequence signature is not present.   
     
     
         17 . A high density nucleic acid array comprising sequence signatures from the TGF-beta gene family.