User:Nzimm: Difference between revisions
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| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA1 Ada1] (aka HFI1, SUP110, SRM12, GAN1) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA1 Ada1] (aka HFI1, SUP110, SRM12, GAN1) | ||
| 1.467 kb/489 aa, Chr. XVI, <br>viable | | 1.467 kb/489 aa, Chr. XVI, <br>viable | ||
| | | "Histone H2A Functional Interactor"; Adapter protein involved in structural integrity of SAGA | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA2 Ada2] (aka SWI8) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA2 Ada2] (aka SWI8) | ||
| 1.305 kb/434aa, Chr. IV, <br>viable | | 1.305 kb/434aa, Chr. IV, <br>viable | ||
| | | "transcriptional ADAptor"; Transcription coactivator | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA3 Ada3](aka NGG1, SWI7) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ADA3 Ada3](aka NGG1, SWI7) | ||
| 2.109 kb/702aa, Chr. IV, <br>viable | | 2.109 kb/702aa, Chr. IV, <br>viable | ||
| | | Transcriptional regulator involved in glucose repression of Gal4p-regulated genes | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=GCN5 Gcn5] (aka ADA4, SWI9) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=GCN5 Gcn5] (aka ADA4, SWI9) | ||
| 1.32 kb/439aa, Chr. VII, <br>viable | | 1.32 kb/439aa, Chr. VII, <br>viable | ||
| | | "General Control Nonderepressible"; Histone acetyltransferase, acetylates N-terminal lysines on histones H2B and H3; this is the catalytic subunit | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Ada5 Ada5] (aka SPT20) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Ada5 Ada5] (aka SPT20) | ||
| 1.815 kb/604aa, Chr. XV, <br>viable | | 1.815 kb/604aa, Chr. XV, <br>viable | ||
| | | "SuPpressor of Ty"; involved in maintaining the integrity of the SAGA transcriptional regulatory complex | ||
|} | |} | ||
{| border="1" | {| border="1" | ||
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| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=spt3 Spt3] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=spt3 Spt3] | ||
| 1.014 kb/337aa, Chr. IV, <br> viable | | 1.014 kb/337aa, Chr. IV, <br> viable | ||
| | | interacts with Spt15p to activate transcription of some RNA polymerase II-dependent genes, also functions to inhibit transcription at some promoters | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Spt7 Spt7](aka GIT2) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Spt7 Spt7](aka GIT2) | ||
| 3.999 kb/1332aa, Chr. II, <br> viable | | 3.999 kb/1332aa, Chr. II, <br> viable | ||
| | | involved in proper assembly of the complex; also present as a C-terminally truncated form in the SLIK/SALSA transcriptional regulatory complex | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Spt8 Spt8] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Spt8 Spt8] | ||
| 1.809 kb/602aa, Chr. XII, <br> viable | | 1.809 kb/602aa, Chr. XII, <br> viable | ||
| | | required for SAGA-mediated inhibition at some promoters; not present in SLIK/SALSA complex | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=SPT20 Spt20] (aka Ada5) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=SPT20 Spt20] (aka Ada5) | ||
| 1.815 kb/604aa, Chr. XV, <br> viable | | 1.815 kb/604aa, Chr. XV, <br> viable | ||
| | | maintains integrity of complex | ||
|} | |} | ||
{| border="1" | {| border="1" | ||
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| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF5 TAF5] (aka TAF90) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF5 TAF5] (aka TAF90) | ||
| 2.397 kb/798aa, Chr. II, <font color = red>inviable</font color> | | 2.397 kb/798aa, Chr. II, <font color = red>inviable</font color> | ||
| | | "TATA binding protein-Associated Factor"; RNA polymerase II transcription initiation and in chromatin modification (90 kDa subunit) | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF6 TAF6] (aka TAF60) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF6 TAF6] (aka TAF60) | ||
| 1.551 kb/516aa, Chr. VII, <font color = red>inviable</font color> | | 1.551 kb/516aa, Chr. VII, <font color = red>inviable</font color> | ||
| | | 60 kDa subunit with same function as above; similar to Histone H4 | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF9 TAF9] (aka TAF17) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF9 TAF9] (aka TAF17) | ||
| 0.474 kb/157aa, Chr. XIII, <font color = red>inviable</font color> | | 0.474 kb/157aa, Chr. XIII, <font color = red>inviable</font color> | ||
| | | 17 kDa subunit; similar to Histone H3 | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF10 TAF10] (aka TAF23, TAF25) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF10 TAF10] (aka TAF23, TAF25) | ||
| 0.621 kb/206aa, Chr. IV, <font color = red>inviable</font color> | | 0.621 kb/206aa, Chr. IV, <font color = red>inviable</font color> | ||
| | | 145 kDa subunit | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF12 TAF12](aka TAF61, TAF68) | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TAF12 TAF12](aka TAF61, TAF68) | ||
| 1.620 kb/539aa, Chr. IV, <font color = red>inviable</font color> | | 1.620 kb/539aa, Chr. IV, <font color = red>inviable</font color> | ||
| | | 61/68 kDa subunit; similar to Histone H2A | ||
|} | |} | ||
{| border="1" | {| border="1" | ||
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| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TRA1 Tra1] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=TRA1 Tra1] | ||
| 11.235 kb/3744aa, Chr. VIII, <font color = red>inviable</font color> | | 11.235 kb/3744aa, Chr. VIII, <font color = red>inviable</font color> | ||
| | | interacts with acidic activators (e.g., Gal4p) which leads to transcription activation; similar to human TRRAP, which is a cofactor for c-Myc mediated oncogenic transformation | ||
|} | |} | ||
{| border="1" | {| border="1" | ||
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| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf73 Sgf73] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf73 Sgf73] | ||
| 1.974 kb/657aa, Chr. VII , <br> viable | | 1.974 kb/657aa, Chr. VII , <br> viable | ||
| | | "SaGa associated Factor"; formation of the preinitiation complex assembly at promoters (73 kDa subunit) | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf29 Sgf29] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf29 Sgf29] | ||
| 0.779 kb/259aa, Chr. III, <br> viable | | 0.779 kb/259aa, Chr. III, <br> viable | ||
| | | 29 kDa subunit of SAGA histone acetyltransferase complex | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf11 Sgf11] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=sgf11 Sgf11] | ||
| 0.3 kb/99aa, Chr.XVI, <br> viable | | 0.3 kb/99aa, Chr.XVI, <br> viable | ||
| | | Integral subunit of SAGA histone acetyltransferase complex, regulates transcription of a subset of SAGA-regulated genes, required for the Ubp8p association with SAGA and for H2B deubiquitylation (11 kDa) | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ubp8 Ubp8] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=ubp8 Ubp8] | ||
| 1.416 kb/471aa, Chr. XIII, <br> viable | | 1.416 kb/471aa, Chr. XIII, <br> viable | ||
| | | Ubiquitin-specific protease - required for SAGA-mediated deubiquitination of histone H2B | ||
|- | |- | ||
| [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Sus1 Sus1] | | [http://db.yeastgenome.org/cgi-bin/locus.pl?locus=Sus1 Sus1] | ||
| gene with intron, Chr. II, <br> viable | | gene with intron, Chr. II, <br> viable | ||
| | | "Sl gene Upstream of ySa1" mRNA export coupled transcription activation; component of the SAGA histone acetylase complex | ||
|} | |} |
Revision as of 11:07, 3 April 2007
Neil Zimmerman
Bio
- MIT Class of 2009
- Major: Biological Engineering
- Minors: Management and Music
- Varsity Soccer
- Wind Ensemble (Clarinet)
20.109
M13K07 Genome Engineering Plans:
Gene | Plans |
---|---|
I | Change # of protein copies expressed to experiment with different sized channels |
II | Modify residues to allow deactivation of p2 under certain conditions so that replication of + strand can be regulated |
III | Insert myc to allow detection with an antibody |
IV | Change # of protein copies expressed to experiment with different sized channels |
V | Add fluorescent tag to monitor levels of p5-ssDNA complex |
VI | Modify residues to help p3 bind more effectively to the ToIA protein on the E. coli F pilus |
VII | Minimize the bulk of the protein to allow more room for modifications on p9 |
VIII | Insert myc (as in III) or another tag to serve as a “hook” for attaching constructs to M13 |
IX | Add residues to N-terminus to present on the outside of the phage coat |
X | Add sensitivity to different stimulus than that of p2 in order to regulate replication of + strand in another fashion |
XI | Modify residues to allow proteins other than p8 to embed in the membrane and serve as the phage filament coat |
M13K07 Genome Refactoring:
In refactoring the M13K07 genome between the HpaI site in gene II and the BamHI site in gene III, I separated all overlapping elements across the region by copying overlapping sequences and placing the elements adjacent to one another on the refactored sequence. As a result, the refactored region is 1,000 bp larger than the original sequence. I also added annotation for & refactored the open reading frames of genes 2, 5, 7, and 10, which are not annotated in the M1307 part in the registry. I inserted the sequence encoding the myc protein into g3. I intend to separate each element with two unique restriction sites, so that the elements can be isolated & manipulated more easily. I initially proposed to add a fluorescent tag like GFP to g5, but seeing as how GFP is nearly 3 times the size of the protein product of g5, I chose to leave g5 unaltered. Below is a table describing the overlaps that were refactored.
Overlapping elements | Action |
---|---|
rbs g7 & g5 ORF | copied overlapping sequence & placed rbs g7 adjacent to the 3' end of the g5 ORF |
promoter g8 & rbs g9 | copied overlapping sequence & placed rbs g9 adjacent to the 3' end of promoter g8 |
promoter g3 & g8 ORF | copied overlapping sqeuence & placed promoter g3 adjacent to the 3' end of g8 ORF |
g10 ORF, g10 rbs, g10 promoter, g5 promoter, & g2 ORF | copied g2 ORF & placed adjacent to 5' end of other (non-overlapping) elements |
inserted the sequence encoding the myc protein into the BamHI site of g3 |
Current Courses
Other
Email: nzimm (at) mit (dot) edu
SAGA subunits, S. cerevisiae
Ada subunits | size,chromosome,null p-type | notes |
---|---|---|
Ada1 (aka HFI1, SUP110, SRM12, GAN1) | 1.467 kb/489 aa, Chr. XVI, viable |
"Histone H2A Functional Interactor"; Adapter protein involved in structural integrity of SAGA |
Ada2 (aka SWI8) | 1.305 kb/434aa, Chr. IV, viable |
"transcriptional ADAptor"; Transcription coactivator |
Ada3(aka NGG1, SWI7) | 2.109 kb/702aa, Chr. IV, viable |
Transcriptional regulator involved in glucose repression of Gal4p-regulated genes |
Gcn5 (aka ADA4, SWI9) | 1.32 kb/439aa, Chr. VII, viable |
"General Control Nonderepressible"; Histone acetyltransferase, acetylates N-terminal lysines on histones H2B and H3; this is the catalytic subunit |
Ada5 (aka SPT20) | 1.815 kb/604aa, Chr. XV, viable |
"SuPpressor of Ty"; involved in maintaining the integrity of the SAGA transcriptional regulatory complex |
Spt subunits | size, chromosome, null p-type | notes |
---|---|---|
Spt3 | 1.014 kb/337aa, Chr. IV, viable |
interacts with Spt15p to activate transcription of some RNA polymerase II-dependent genes, also functions to inhibit transcription at some promoters |
Spt7(aka GIT2) | 3.999 kb/1332aa, Chr. II, viable |
involved in proper assembly of the complex; also present as a C-terminally truncated form in the SLIK/SALSA transcriptional regulatory complex |
Spt8 | 1.809 kb/602aa, Chr. XII, viable |
required for SAGA-mediated inhibition at some promoters; not present in SLIK/SALSA complex |
Spt20 (aka Ada5) | 1.815 kb/604aa, Chr. XV, viable |
maintains integrity of complex |
TAF subunits | size, chromosome, null p-type | notes |
---|---|---|
TAF5 (aka TAF90) | 2.397 kb/798aa, Chr. II, inviable | "TATA binding protein-Associated Factor"; RNA polymerase II transcription initiation and in chromatin modification (90 kDa subunit) |
TAF6 (aka TAF60) | 1.551 kb/516aa, Chr. VII, inviable | 60 kDa subunit with same function as above; similar to Histone H4 |
TAF9 (aka TAF17) | 0.474 kb/157aa, Chr. XIII, inviable | 17 kDa subunit; similar to Histone H3 |
TAF10 (aka TAF23, TAF25) | 0.621 kb/206aa, Chr. IV, inviable | 145 kDa subunit |
TAF12(aka TAF61, TAF68) | 1.620 kb/539aa, Chr. IV, inviable | 61/68 kDa subunit; similar to Histone H2A |
Tra1 subunit | size, chromosome, null p-type | notes |
---|---|---|
Tra1 | 11.235 kb/3744aa, Chr. VIII, inviable | interacts with acidic activators (e.g., Gal4p) which leads to transcription activation; similar to human TRRAP, which is a cofactor for c-Myc mediated oncogenic transformation |
other subunits | size, chromosome, null p-type | notes |
---|---|---|
Sgf73 | 1.974 kb/657aa, Chr. VII , viable |
"SaGa associated Factor"; formation of the preinitiation complex assembly at promoters (73 kDa subunit) |
Sgf29 | 0.779 kb/259aa, Chr. III, viable |
29 kDa subunit of SAGA histone acetyltransferase complex |
Sgf11 | 0.3 kb/99aa, Chr.XVI, viable |
Integral subunit of SAGA histone acetyltransferase complex, regulates transcription of a subset of SAGA-regulated genes, required for the Ubp8p association with SAGA and for H2B deubiquitylation (11 kDa) |
Ubp8 | 1.416 kb/471aa, Chr. XIII, viable |
Ubiquitin-specific protease - required for SAGA-mediated deubiquitination of histone H2B |
Sus1 | gene with intron, Chr. II, viable |
"Sl gene Upstream of ySa1" mRNA export coupled transcription activation; component of the SAGA histone acetylase complex |