User:Samiye Yaman/Notebook/Lab 571/2011/09/27

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Entry title

Objective

Description

-The protein was placed in a a beaker full of water in order to defrost.

-The protein was made up of maltose binding protein(MBP), Mxe GyrA Intein and Chitin binding protein. Protein sequence of all three was found and calculated by using protein calculator (http://www.scripps.edu/~cdputnam/protcalc.html)

-PI of the protein was calculated as 7.32 and the mass of the protein was 1348987.79824 Da. The protein sequence is

MBP

MGNHAGKREL NAEKASTNSE TNRGESEKKR NLGELSRTTS EDNEVFGEAD ANQNNGTSSQ DTAVTDSKRT ADPKNAWQDA HPADPGSRPH LIRLFSRDAP GREDNTFKDR PSESDELQTI QEDSAATSES LDVMASQKRP SQRHGSKYLA TASTMDHARH GFLPRHRDTG ILDSIGRFFG GDRGAPKRGS GKDSHHPART AHYGSLPQKS HGRTQDENPV VHFFKNIVTP RTPPPSQGKG RGLSLSRFSW GAEGQRPGFG YGGRASDYKS AHKGFKGVDA QGTLSKIFKL GGRDSRSGSP


CBD

MKLKLTSKLE RLRTSKKGIG ACVLAAGMTA ITMAPQSAYA HGFVEKPSSR AALCSQNYGV LNLNCGNVMY EPQSLEAPKG FPDSGPIDGK IASAGGLFGG ILDQQTSNRW FKNTIKGGVN TFTWKYTAAH STSKWHYYIT KKGWDPNKPL TRAELEPIGT VKHDGSAASN NLTHTINVPT DRNGYHVILA VWDVADTSNA FYNVVDVNLV NNETPDTVAP SQPTELNASK VSANSVEITW KASTDNIGVK EYQVLRNGEV IDTVPGTTFI DKKLKADTEY TYTIKALDSA GNISKESEKL KVKTTHTIPD IEAPTQPKGL HSMGTTSTTV DLMWSPSEDN VGVDHYIVYR ESAGVMNKIG TAANTSFMDK DLKANTSYNY VVTAVDLAGN ESSRSDVLNV TTKSENSAYE KWDARKAYTK GDRVVHEGKV YEAVQNHQGN GDSNWIFALS LWKPVLNK


Gyr A

MTDTTLPPEG EAHDRIEPVD IQQEMQRSYI DYAMSVIVGR ALPEVRDGLK PVHRRVLYAM YDSGFRPDRS HAKSARSVAE TMGNYHPHGD ASIYDTLVRM AQPWSLRYPL VDGQGNFGSP GNDPPAAMRY TEARLTPLAM EMLREIDEET VDFIPNYDGR VQEPTVLPSR FPNLLANGSG GIAVGMATNI PPHNLGELAE AVYWCLENYE ADEEATCEAV MERVKGPDFP TSGLIVGTQG IEDTYKTGRG SIKMRGVVEI EEDSRGRTSI VITELPYQVN HDNFITSIAE QVRDGKLAGI SNIEDQSSDR VGLRIVVELK RDAVAKVVLN NLYKHTQLQT SFGANMLSIV DGVPRTLRLD QLIRLYVDHQ LDVIVRRTRY RLRKANERAH ILRGLVKALD ALDEVIALIL VALATGRAGD ITLDDLHRPE TFFKTFTQTG CTRVQFFEYK QSTLSFGEGT RTGCLSMNS


-The protein samples were distributed equally to 4 plastic graduated tubes for sonification. Each sample was sonificated for 30 sec and waited for 30 sec. This sonification was repeated three times for each sample.

-After balancing the mass of each samples with tubes, proteins were placed into the centrifuge at 15000rmp, 4 degree celcius for an hour.

-By using 24.228 g of tris and 4 L of water, a buffer with 9.01 ph was prepared. Same procedure was repeated to form same buffer. (4L of two buffers placed in different buckets.) Ph of Tris was around 10. By adding NaOH and HCl solution, the ph was able to set at 9.01.

- After the centrifuge, the protein solution from each plastic tube were placed into two dialysis tube.

-Each dialysis tube was placed into two different buckets.

-Into two different 1 ml flask 2 different buffers were prepared. One of the buffers contained 6.057 g of tris and 1 l of water. The other buffer contained again 6.057 g of Tris, 1 l of water and 58.44 g of NaCl.

-By adding NaOH and HCL, the pH of each buffers were arranged to 9.Both of the buffers had a final ph Of 8.98.

-Each of the flask with buffers covered with aluminum foil.

Data

  • Add data and results here...

PI of the protein was calculated as 7.32 and the mass of the protein was 1348987.79824 Da. The protein sequence is

MBP

MGNHAGKREL NAEKASTNSE TNRGESEKKR NLGELSRTTS EDNEVFGEAD ANQNNGTSSQ DTAVTDSKRT ADPKNAWQDA HPADPGSRPH LIRLFSRDAP GREDNTFKDR PSESDELQTI QEDSAATSES LDVMASQKRP SQRHGSKYLA TASTMDHARH GFLPRHRDTG ILDSIGRFFG GDRGAPKRGS GKDSHHPART AHYGSLPQKS HGRTQDENPV VHFFKNIVTP RTPPPSQGKG RGLSLSRFSW GAEGQRPGFG YGGRASDYKS AHKGFKGVDA QGTLSKIFKL GGRDSRSGSP


CBD

MKLKLTSKLE RLRTSKKGIG ACVLAAGMTA ITMAPQSAYA HGFVEKPSSR AALCSQNYGV LNLNCGNVMY EPQSLEAPKG FPDSGPIDGK IASAGGLFGG ILDQQTSNRW FKNTIKGGVN TFTWKYTAAH STSKWHYYIT KKGWDPNKPL TRAELEPIGT VKHDGSAASN NLTHTINVPT DRNGYHVILA VWDVADTSNA FYNVVDVNLV NNETPDTVAP SQPTELNASK VSANSVEITW KASTDNIGVK EYQVLRNGEV IDTVPGTTFI DKKLKADTEY TYTIKALDSA GNISKESEKL KVKTTHTIPD IEAPTQPKGL HSMGTTSTTV DLMWSPSEDN VGVDHYIVYR ESAGVMNKIG TAANTSFMDK DLKANTSYNY VVTAVDLAGN ESSRSDVLNV TTKSENSAYE KWDARKAYTK GDRVVHEGKV YEAVQNHQGN GDSNWIFALS LWKPVLNK


Gyr A

MTDTTLPPEG EAHDRIEPVD IQQEMQRSYI DYAMSVIVGR ALPEVRDGLK PVHRRVLYAM YDSGFRPDRS HAKSARSVAE TMGNYHPHGD ASIYDTLVRM AQPWSLRYPL VDGQGNFGSP GNDPPAAMRY TEARLTPLAM EMLREIDEET VDFIPNYDGR VQEPTVLPSR FPNLLANGSG GIAVGMATNI PPHNLGELAE AVYWCLENYE ADEEATCEAV MERVKGPDFP TSGLIVGTQG IEDTYKTGRG SIKMRGVVEI EEDSRGRTSI VITELPYQVN HDNFITSIAE QVRDGKLAGI SNIEDQSSDR VGLRIVVELK RDAVAKVVLN NLYKHTQLQT SFGANMLSIV DGVPRTLRLD QLIRLYVDHQ LDVIVRRTRY RLRKANERAH ILRGLVKALD ALDEVIALIL VALATGRAGD ITLDDLHRPE TFFKTFTQTG CTRVQFFEYK QSTLSFGEGT RTGCLSMNS

Notes

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