BLAST  Project (40 pts) - due Oct. 6th

Assignments:  In addition to writing on exams and homework, there will be two special assignments.    

 

To earn an excellent grade on your report, you must:

·                     communicate clearly;   make it interesting and informative to the reader; 

·                     answer all listed questions, with your thinking clearly explained;

·                     turn the report in on time (late penalty - 4 pts/ first day + 2 pts/additional day; no credit after 10 days)

·                      format for your reports:

o         Place your name, UTeID, class, date, seq #, 20-mer seq., name of protein, complete FASTA seq. properly formatted, etc.  information on the cover page

o        use one and one-half spacing for the body of your text

o        use single spacing for figure captions and references

o        references: include at least 4 primary references (see below) in J Biol Chem style - place a number within the text (1) and then provide a list of references at the end, single spaced.  e.g., Borst, P., and Elferink, R. O. (2002) Annu. Rev. Biochem. 71, 537?592 .

o        make sure that Figures included have a figure legend and Tables have a title or heading 

              be neat, with few or no spelling or grammatical errors.

Assignment 1. You will be assigned a 20-amino acid sequence from the list below.  Each sequence is a fragment of an actual protein.  Use your sequence to do the following steps and then answer the questions below for your report:  

(a) Do a first BLAST run using your 20 a.a. sequence.  What is your protein? What is the complete amino acid sequence of the protein?  

(b) Now using the entire a.a. sequence, do a second BLAST run to find related sequences.  Carefully select a subset of the related sequences and do a Multiple Sequence Alignment using a program like CLUSTAL W.   Choose related proteins from at least five to eight other diverse organisms, if possible.  Using relatively diverse organisms (archaeal, bacterial, and eukaryotic) will make it easier to identify the most conserved amino acids within the protein.   Prepare a figure or table showing a sequence alignment of your protein and identify the conserved amino acids.  

(c)  Which amino acids (be specific, aa #) do you think are most important for preserving the structure of the protein?  Why?  Which amino acids do you think are most important for the function of the protein?  Why?   

(d) You should find that a structure is known for your protein.  What is the PDB code?

(e)  Using PyMOL (or a similiar program) make an original figure illustrating your protein.  

(f)  In approximately one-half page, briefly describe the function of your protein.  

REPORT / Grading - 40 pts total

1) Cover page - your name, the date and class, your sequence number and given 20 a.a. sequence, the name and source of your protein and the complete protein sequence in FASTA format -  5 pts

3)  Table showing your multiple sequence alignment using 5 selected BLAST hits - 15 pts

4) Original PyMOL or similar figure - 10 pts, and

5) Half-page functional summary of your protein - 10 pts.  Tell a clear, story with a  minimum of typos / errors and include four (4) primary references to go with your report (see above).

Regarding plagiarism:  Please be aware that plagiarism is considered to be ethically unacceptable. Here is a one suggestion as to how to avoid plagiarism: While reading the sources of information that you plan to use in preparing your assignment, take notes on the content, using your own words.  While preparing your paper, refer to your notes, rather than the original source.  If you feel the need to use a phrase from a source, be sure to put the phrase in quotes, and reference the source.  In these days of the internet, it is very easy to find material that can be easily be plagiarized. Be warned, however, that the existence of the internet makes it much easier than ever before to detect plagiarism.  If you plagiarize in writing assignments, you run a high risk of getting caught, and a high risk of failing the course.

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Select your 20 amino acid sequence for the first writing assignment by the day of your birth from the list below  (e.g. if you were born on September  11, 2001 - your sequence is VNFKIRHNIE DGSVQLADHY

             Sequences for CH370  2011  -  BLAST Writing Assignment

1)  LLTRTGNPIF APQWFTNGLP    2) PIRLEGLVLT HCNFSSYEPE 

3)  VELGANWVEG VNGGKMNPIW    4) TDDLEKVCNE VVDTCLYKGS

5)  FAMLSLGTKA DTHDEILEGL    6) SADYVMVSAS LGVLQSDLIQ 

7)  HLKEGLVAII DLAVDRLHCS    8) MPMSERSVGG SGAGGGGDAL

9)  NASCHGTNGT VHAGPRDTGD   10) HQLKNNPSSR RHITMLWNPD           

11) VNFKIRHNIE DGSVQLADHY   12) STEEYNKWKF YNRPNILEVL

13) SQPHRPALSF RDLFSMATEE   14) LSRVFGDLRE GKRILREMEI

15) GRRFLLRRKF IPDPGGTNLM   16) YTRVLPSVPQ DCPTPMGTKG

17) LRRSPAPNPP AAASPTTTEA   18) DTIENVKAKI QDKEGIPPDQ     

19) SYMGFEVVRP DHPALPPLDN   20) RYLCKKQGTK KTLQKIGAAC     

21) LGGYAPGNAY LDGLAQQRRS   22) TFKQECKIKY SERVYDACMD

23) QNNFVHDCVN ITVKEHTVTT   24) VGKKREFVER LTSVAAEIYG

25) GGNGIVGAQV PLGAGIALAC   26) PSGNIGANGV AIFESVHGTA 

27) RRSASLHLPK LSITGTYDLK   28) TQGQIYGIKV DIRDAYGNVK      

29) QNRSYSKLLC GLLAERLRIS   30) RDNMSVILIC FPNAPKVSPE  

31) LGKKNRSLNG EKVDQVDYLL    

 

Every sequence should have a perfect match with a real protein, but that protein may not have a structure that has been determined.  However, if that is the case you should be able to find a closely related protein that does have a structure available.