Difference between revisions of "Part:BBa K1796013"
(→‎Parameter of Protein)
 
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==Parameter of Protein==
 
==Parameter of Protein==
−
<p> Number of amino acids: 261 </p>
+
<p> Number of amino acids: 253 </p>
 
<p> </p>
 
<p> </p>
−
<p> Molecular weight: 28332.5 </p>
+
<p> Molecular weight: 27608.7 </p>
 
<p> </p>
 
<p> </p>
−
<p> Theoretical pI: 5.51 </p>
+
<p> Theoretical pI: 5.29 </p>
 
<p> </p>
 
<p> </p>
 
<p> Amino acid composition:  </p>
 
<p> Amino acid composition:  </p>
 
<p> </p>
 
<p> </p>
−
<p> Ala (A)  26      10.0% </p>
+
<p> Ala (A)  25      9.9% </p>
 
<p> </p>
 
<p> </p>
−
<p> Arg (R)  18       6.9% </p>
+
<p> Arg (R)  17       6.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Asn (N)  7      2.7% </p>
+
<p> Asn (N)  7      2.8% </p>
 
<p> </p>
 
<p> </p>
−
<p> Asp (D)  12      4.6% </p>
+
<p> Asp (D)  12      4.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Cys (C)  8      3.1% </p>
+
<p> Cys (C)  8      3.2% </p>
 
<p> </p>
 
<p> </p>
−
<p> Gln (Q)  7      2.7% </p>
+
<p> Gln (Q)  7      2.8% </p>
 
<p> </p>
 
<p> </p>
−
<p> Glu (E)  20      7.7% </p>
+
<p> Glu (E)  20      7.9% </p>
 
<p> </p>
 
<p> </p>
−
<p> Gly (G)  31     11.9% </p>
+
<p> Gly (G)  27     10.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> His (H)  8       3.1% </p>
+
<p> His (H)  7       2.8% </p>
 
<p> </p>
 
<p> </p>
−
<p> Ile (I)  13      5.0% </p>
+
<p> Ile (I)  13      5.1% </p>
 
<p> </p>
 
<p> </p>
−
<p> Leu (L)  27      10.3% </p>
+
<p> Leu (L)  27      10.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Lys (K)  6      2.3% </p>
+
<p> Lys (K)  6      2.4% </p>
 
<p> </p>
 
<p> </p>
−
<p> Met (M)  13       5.0% </p>
+
<p> Met (M)  12       4.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Phe (F)  3      1.1% </p>
+
<p> Phe (F)  3      1.2% </p>
 
<p> </p>
 
<p> </p>
−
<p> Pro (P)  12      4.6% </p>
+
<p> Pro (P)  12      4.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Ser (S)  10      3.8% </p>
+
<p> Ser (S)  10      4.0% </p>
 
<p> </p>
 
<p> </p>
−
<p> Thr (T)  12      4.6% </p>
+
<p> Thr (T)  12      4.7% </p>
 
<p> </p>
 
<p> </p>
−
<p> Trp (W)  3      1.1% </p>
+
<p> Trp (W)  3      1.2% </p>
 
<p> </p>
 
<p> </p>
−
<p> Tyr (Y)  9      3.4% </p>
+
<p> Tyr (Y)  9      3.6% </p>
 
<p> </p>
 
<p> </p>
−
<p> Val (V)  16      6.1% </p>
+
<p> Val (V)  16      6.3% </p>
 
<p> </p>
 
<p> </p>
 
<p> Pyl (O)  0      0.0% </p>
 
<p> Pyl (O)  0      0.0% </p>
Line 79: Line 79:
 
<p> </p>
 
<p> </p>
 
<p> Total number of negatively charged residues (Asp + Glu): 32 </p>
 
<p> Total number of negatively charged residues (Asp + Glu): 32 </p>
−
<p> Total number of positively charged residues (Arg + Lys): 24 </p>
+
<p> Total number of positively charged residues (Arg + Lys): 23 </p>
 
<p> </p>
 
<p> </p>
−
<p> Atomic composition:Carbon      C              1231 </p>
+
<p> Atomic composition:Carbon      C              1203 </p>
−
<p> Hydrogen    H        1964 </p>
+
<p> Hydrogen    H        1919 </p>
−
<p> Nitrogen    N          354 </p>
+
<p> Nitrogen    N          341 </p>
−
<p> Oxygen      O          371 </p>
+
<p> Oxygen      O          363 </p>
−
<p> Sulfur      S          21 </p>
+
<p> Sulfur      S          20 </p>
 
<p> </p>
 
<p> </p>
−
<p> Formula: C1231H1964N354O371S21Total number of atoms: 3941 </p>
+
<p> Formula: C1203H1919N341O363S20Total number of atoms: 3846 </p>
 
<p> </p>
 
<p> </p>
 
<p> Extinction coefficients:Extinction coefficients are in units of  M-1 cm-1, at 280 nm measured in water. </p>
 
<p> Extinction coefficients:Extinction coefficients are in units of  M-1 cm-1, at 280 nm measured in water. </p>
 
<p> </p>
 
<p> </p>
 
<p> Ext. coefficient    30410 </p>
 
<p> Ext. coefficient    30410 </p>
−
<p> Abs 0.1% (=1 g/l)  1.073, assuming all pairs of Cys residues form cystines </p>
+
<p> Abs 0.1% (=1 g/l)  1.101, assuming all pairs of Cys residues form cystines </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> Ext. coefficient    29910 </p>
 
<p> Ext. coefficient    29910 </p>
−
<p> Abs 0.1% (=1 g/l)  1.056, assuming all Cys residues are reduced </p>
+
<p> Abs 0.1% (=1 g/l)  1.083, assuming all Cys residues are reduced </p>
 
<p> </p>
 
<p> </p>
−
<p> Estimated half-life:The N-terminal of the sequence considered is A (Ala). </p>
+
<p> Estimated half-life:The N-terminal of the sequence considered is V (Val). </p>
 
<p> </p>
 
<p> </p>
−
<p> The estimated half-life is: 4.4 hours (mammalian reticulocytes, in vitro). </p>
+
<p> The estimated half-life is: 100 hours (mammalian reticulocytes, in vitro). </p>
 
<p>                             >20 hours (yeast, in vivo). </p>
 
<p>                             >20 hours (yeast, in vivo). </p>
 
<p>                             >10 hours (Escherichia coli, in vivo). </p>
 
<p>                             >10 hours (Escherichia coli, in vivo). </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
−
<p> Instability index:The instability index (II) is computed to be 43.97 </p>
+
<p> Instability index:The instability index (II) is computed to be 44.33 </p>
 
<p> This classifies the protein as unstable. </p>
 
<p> This classifies the protein as unstable. </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
 
<p> </p>
−
<p> Aliphatic index: 87.51 </p>
+
<p> Aliphatic index: 89.88 </p>
 +
<p> </p>
 +
<p> Grand average of hydropathicity (GRAVY): -0.077 </p>
 +
<p> </p>
 +
<p> </p>
 
<p> </p>
 
<p> </p>
−
<p> Grand average of hydropathicity (GRAVY): -0.097 </p>
 

Latest revision as of 01:50, 19 September 2015

hesA promoted from Paenibacillus sp. WLY78

Function has not so far been determined , but Deletion analysis indicates that in addition to the core nif genes, hesA plays an important role in nitrogen fixation and is responsive to the availability of molybdenum. HesA a member of the ThiF-MoeB-HesA family and contains an N-terminal nucleotide binding domain and a C-terminal MoeZ/MoeB-like domain. The ThiF-MoeB-HesA family engage in an ATP-dependent process that activates the C-terminus of partner ubiquitin-like proteins by forming an acyl adenylate complex that facilitates sulfur transfer.

First, we promoted it ourselves（done by Nan Wang and Nannan Xie).Then we sent the sequences to synthesis, but unfortunately, striction enzyme cut site was involved after they promoted it again. But we were not informed of the error in promotion.After several times failed expriments, we found the problem.We tackled them over the matter, mistakes were corrected, but the correct genes can't arrive on time,it can only arrive after the deadline. Sequence submit is promoted by the synthesis company, containing a PstI in the gene

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 143
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 143
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 143
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 143
  • 1000
    COMPATIBLE WITH RFC[1000]


Parameter of Protein

Number of amino acids: 253

Molecular weight: 27608.7

Theoretical pI: 5.29

Amino acid composition: 

Ala (A) 25 9.9%

Arg (R) 17 6.7%

Asn (N) 7 2.8%

Asp (D) 12 4.7%

Cys (C) 8 3.2%

Gln (Q) 7 2.8%

Glu (E) 20 7.9%

Gly (G) 27 10.7%

His (H) 7 2.8%

Ile (I) 13 5.1%

Leu (L) 27 10.7%

Lys (K) 6 2.4%

Met (M) 12 4.7%

Phe (F) 3 1.2%

Pro (P) 12 4.7%

Ser (S) 10 4.0%

Thr (T) 12 4.7%

Trp (W) 3 1.2%

Tyr (Y) 9 3.6%

Val (V) 16 6.3%

Pyl (O) 0 0.0%

Sec (U) 0 0.0%

(B) 0 0.0%

(Z) 0 0.0%

(X) 0 0.0%

Total number of negatively charged residues (Asp + Glu): 32

Total number of positively charged residues (Arg + Lys): 23

Atomic composition:Carbon C 1203

Hydrogen H 1919

Nitrogen N 341

Oxygen O 363

Sulfur S 20

Formula: C1203H1919N341O363S20Total number of atoms: 3846

Extinction coefficients:Extinction coefficients are in units of M-1 cm-1, at 280 nm measured in water.

Ext. coefficient 30410

Abs 0.1% (=1 g/l) 1.101, assuming all pairs of Cys residues form cystines

Ext. coefficient 29910

Abs 0.1% (=1 g/l) 1.083, assuming all Cys residues are reduced

Estimated half-life:The N-terminal of the sequence considered is V (Val).

The estimated half-life is: 100 hours (mammalian reticulocytes, in vitro).

>20 hours (yeast, in vivo).

>10 hours (Escherichia coli, in vivo).

Instability index:The instability index (II) is computed to be 44.33

This classifies the protein as unstable.

Aliphatic index: 89.88

Grand average of hydropathicity (GRAVY): -0.077