Open Reading Frames

In bioinformatics we look for gene-coding sequences or what we call open reading frames(ORF), entrez has a tool called ORF finder (now you know why I like entrez :-). 
http://www.ncbi.nlm.nih.gov/projects/gorf/ Let's use this tool to find out what regions will code for a gene in this sequence.

For your Classwork
ACTTTGCAGGCAGCGGCGGCCGGGGCGGAGCGGGATCGAGCCCTCGCCGCGGCCTGCCAGTCATGGGCCCGCGCCGCCGCCGCCGCCTGCCTCCCGGGCCACGCGGGCCGTGAGCGCCATGGCCGTAGCCCCCGCGGGCGGCCAGCACGCGCCAGCGCTGGAGGCCCTGCTCGGGGCGGGCGCGTTGCGGCTGCTCGACTCCTCGCAGATCGTCATCATCTCCACCGCGCCCGATGTCGGCGCCCCGCAGCTCCCCGCCGCGCCGCCCACTGGCCCTCGCGATTCTGACGTGCTGCTCTTCGCCACGCCGCAGGCGCCCCGACCCGCGCCTAGTGCACCGCGCCCGGCTCTCGGCCGCCCGCCGGTGAAACGGAGGCTGGATCTGGAGACTGACCATCAGTACCTCGCTGGTAGCAGTGGGCCATTCCGGGGCAGAGGCCGCCACCCAGGGAAAGGTGTGAAATCTCCGGGGGAGAAGTCACGCTATGAAACCTCACTAAATCTGACCACCAAACGCTTCTTGGAGCTGCTGAGCCGCTCAGCTGACGGTGTCGTTGACCTGAACTGGGCAGCTGAGGTGCTGAAGGTGCAGAAACGGCGCATCTATGACATCACCAATGTCCTGGAGGGCATCCAGCTCATTGCCAAGAAGTCCAAGAATCATATCCAGTGGCTAGGCAGCCACACCATGGTGGGGATTGGTAAGCGGCTTGAAGGCCTGACCCAGGACCTGCAGCAACTGCAGGAGAGTGAGCAGCAGCTGGATCACCTGATGCACATCTGTACCACACAGCTGCAACTGCTTTCGGAGGACTCCGACACCCAGCGCCTGGCCTATGTGACCTGCCAGGACCTTCGCAGCATTGCAGACCCTGCAGAACAGATGGTCATAGTGATCAAGGCCCCTCCTGAGACCCAACTACAAGCTGTGGATTCTTCAGAGACATTTCAGATCTCCCTTAAGAGCAAACAAGGCCCCATTGATGTTTTCCTGTGCCCGGAGGAGAGTGCAGACGGGATTAGCCCTGGGAAGACCTCATGCCAGGAGACATCCTCTGGGGAGGACCGGACTGCAGACTCTGGCCCAGCAGGGCCTCCACCATCACCTCCCTCCACATCCCCAGCCTTGGATCCCAGTCAATCCCTGTTGGGCCTGGAGCAAGAAGCAGTATTGCCACGGATGGGCCACCTGAGGGTCCCTATGGAAGAGGACCAACTGTCACCACTGGTGGCTGCTGACTCACTCCTGGAGCATGTTAAAGAAGACTTCTCTGGGCTCCTCCCTGGGGAGTTCATCAGCCTCTCCCCACCCCACGAGGCCCTTGACTATCACTTTGGTCTCGAGGAGGGTGAGGGCATTAGAGATCTCTTTGACTGTGACTTTGGGGACCTGACCCCTCTGGATTTCTGACAGAAGCCTAGGGATTCAGGGTGTCTGGAGATGCCCACCTGTCTGCAGCTTTGGAGCCTCCTGCCCTGGGCCATCCTTCCTGCCTCATTGGAATAGCACGATCCATACCCTCTGTCCCAATAGCTTCTAGCTCTGGGGTTTGGTTGCTGCCACATTGAGCAGACCAAAATGGGAAGGATGTTGTACAGTGTGTGTGCATGCACCCCACACTGCGCACTGTGTGCCTGGGGTGTGTGTCTGAGTGTGTGTGTGTGTGTGTGTGTGTGAGTGTGTGTGTGTGTGTGTGTGAGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTATGTATGTGTATGTGCACGTGTGCCCGGGAATGAAGGTGAACACATCTGTATGTGTGCTGCAGACACATCCTGGTGTGTCCACATGTGTGCATGGATCCATGTGTGCGCATTGGGGTGGGGGTGGGCTCTAACTGCACTTTTGGTGTCCTTGCTGCAGGGGCCCTGTGAGGCCCAGGGTGGCTGCCTGCTTTCAGAATCCTGTGTGTCAGCCAGGCCGGGTGGTACAGCTTGCCTGGCTGGGTTTGCAGGGCAGCAAGAGCACTGCTTAAAAGTTTTCCGATCGAAGCTTTAATGGAGCGTTTATTTATTTATCGAGGCCTCTGGCAAGCCTGGGGGGATAAGCAAAGGGTGGGGGGCATGGGTGATACCTTAAGTCCCTGTTCTCTGAAGCAAGGGCAGGATCCCTACCCAAGAGTTGCTGAGGCCCAAGCAGTTTATTTATTGGGAAAGGGAGAGGGAGACAGACTGACAGCCATGGATGGGCTGGAGAAACAGTCCCTTTGTACCAGTACTCCAGCCGCATGTATCCAGGGGATCTGAGATGGGGAGGGTACGTGAGGGCCTTGGCTGACTGCGGCCAGGAGGGGTGGGTATGCGTCCTTCCTATGGCTGGAGTGCTCCTCTGCTGTCCTCCCCACCCTCCAGTCTGCACTTTGATTTGTTTCCTAACAGTTCTGTTCCCTCCTGCTTTGATTTTAATAAATGTTTTGATG

1. Find the ORF regions
2. Which region is most probably the gene coding region if this sequence contains only a single gene?
3. What is the length of this most probable gene?
4. The gene will encode a protein molecule. How long will this protein molecule be?
5. How many Methionines are encoded in the gene-containing region?
6. In which position is the stop codon found?
7. What is the name of this most probable gene?
8. How did you determine the name of the best-matched gene name: i.e. what was your e-value, total score, number of gaps, %identity.

EXERCISE 1 [ ORF VERSUS GENES]

1.     In our original sequenced DNA, the following ORFs were predicted:

(a)  What is an ORF and how different is it from a gene coding region?                        (2)

(b)  Which ORF is most probably the gene-coding region? Circle it.                   (1)

(c)  How long is this open reading frame?_________________                          (1)

(d)  In which frame was this most probable ORF found?______________        (1)

(e)  Predict the length of the protein that would be coded by this ORF_________(1)

2.     The 3D structure of the resulting gene product looks like this. Describe the protein.

 (7)

Protein Sturctures Databases

The PDB archive contains information about experimentally-determined structures of proteins, nucleic acids, and complex assemblies. As a member of the wwPDB, the RCSB PDB curates and annotates PDB data according to agreed upon standards.

Use the RCSB PDB to perform simple and advanced searches based on annotations relating to sequence, structure and function, and to visualize, download, and analyze molecules.

Choose a protein of your choice according to your current research focus and view the structural components on RCSB http://www.rcsb.org/pdb/home/home.do

Have fun

For your class-work: fully describe the protein structure

Functional Analysis of proteins

Today, let's open expassy and use prosite to look at functional characterization of this protein sequence:

MVQRWLYSTNAKDIAVLYFMLAIFSGMAGTAMSLIIRLELAAPGSQYLHGNSQLFNVLVVGHAVLMIFCAPFRLIYHCIEVLIDKHISVYSINENFTVSFWFWLLVVTYMVFRYVNHMAYPVGANSTGTMACHKSAGVKQPAQGKNCPMARLTNSCKECLGFSLTPSHLGIVIHAYVLEEEVHELTKNESLALSKSWHLEGCTSSNGKLRNTGLSERGNPGDNGVFMVPKFNLNKVRYFSTLSKLNARKEDSLAYLTKINTTDFSELNKLMENNHNKTETINTRILKLMSDIRMLLIAYNKIKSKKGNMSKGSNNITLDGINISYLNKLSKDINTNMFKFSPVRRVEIPKTSGGFRPLSVGNPREKIVQESMRMMLEIIYNNSFSYYSHGFRPNLSCLTAIIQCKNYMQYCNWFIKVDLNKCFDTIPHNMLINVLNERIKDKGFMDLLYKLLRAGYVDKNNNYHNTTLGIPQGSVVSPILCNIFLDKLDKYLENKFENEFNTGNMSNRGRNPIYNSLSSKIYRCKLLSEKLKLIRLRDHYQRNMGSDKSFKRAYFVRYADDIIIGVMGSHNDCKNILNDINNFLKENLGMSINMDKSVIKHSKEGVSFLGYDVKVTPWEKRPYRMIKKGDNFIRVRHHTSLVVNAPIRSIVMKLNKHGYCSHGILGKPRGVGRLIHEEMKTILMHYLAVGRGIMNYYRLATNFTTLRGRITYILFYSCCLTLARKFKLNTVKKVILKFGKVLVDPHSKVSFSIDDFKIRHKMNMTDSNYTPDEILDRYKYMLPRSLSLFSGICQICGSKHDLEVHHVRTLNNAANKIKDDYLLGRMIKMNRKQITICKTCHFKVHQGKYNGPGL

Click on: http://www.expasy.ch/
and open PROSITE
Look at the following:
0. Domain structure of the protein
1. Clustal format(1st 3 sequences)• Retrieve the sequence LOGO from the alignment (for 15 aas)
2. Taxonomic tree view of all Swiss-Prot/TrEMBL entries matching our protein
3. Retrieve a list of all Swiss-Prot/TrEMBL entries matching our protein
4. Scan Swiss-Prot/TrEMBL entries against our protein
5. view ligand binding statistics on our protein
6. Click on sequence ID and retrieve sequence Logo from alignment

For your classwork, here is your sequence
MLDQQTINIIKATVPVLKEHGVTITTTFYKNLFAKHPEVRPLFDMGRQESLEQPKALAMT
VLAAAQNIENLPAILPAVKKIAVKHCQAGVAAAHYPIVGQELLGAIKEVLGDAATDDILD
AWGKAYGVIADVFIQVEADLYAQAVE