Part:BBa_K3970006

CpcA

Usage and Biology

CpcA gene encodes α-phycocyanin, a light-harvesting photosynthetic bile pigment-protein from the phycobiliprotein complex (phycobilisome, PBS). Phycocyanin is the major phycobiliprotein in the PBS rod.

Optimized

Original Sequence (Original Sequence Length: 489bp,GC%: 54.4):

ATGAAAACACCGATTACTGAAGCCATTGCTGCAGCCGATACTCAAGGCCGTTTCTTGAGCAATACTGAACTGCAAGCGGCTGATGGTCGCTTCAAGCGTGCCGTTGCCAGCATGGAA

GCAGCTCGTGCTCTCACCAACAATGCGCAAAGCCTGATCGATGGTGCTGCCCAAGCGGTGTATCAAAAATTCCCCTACACCACCACCATGCAAGGCTCTCAGTATGCATCGACCCCC

GAAGGCAAAGCCAAGTGTGCTCGGGACATCGGCTACTATCTGCGGATGGTGACCTACTGTCTTGTCGCCGGTGGTACCGGCCCAATGGATGAGTACCTGATTGCTGGTTTGGCAGA

AATCAACAGCACCTTTGATCTGTCCCCCAGCTGGTACGTGGAAGCCCTGAAGTACATCAAAGCTAACCATGGGTTGAGCGGCCAGGCAGCGGTGGAAGCCAACGCCTACATCGACT

ACGCCATTAACGCCCTCAGCTAA

Optimized Sequence(Optimized Sequence Length:489bp,GC%:46.22):

ATGAAGACCCCTATCACCGAAGCCATTGCCGCTGCTGACACTCAAGGCCGTTTTTTGTCCAACACTGAATTGCAAGCTGCTGATGGTAGATTCAAGAGAGCTGTTGCTTCCATGGAAG

CTGCCAGAGCTTTGACCAACAACGCCCAATCTTTGATTGATGGTGCTGCACAAGCCGTCTACCAAAAGTTCCCATACACTACCACCATGCAAGGTTCTCAATATGCTTCTACTCCAGAA

GGTAAGGCTAAGTGTGCTAGAGATATCGGTTACTACTTGAGAATGGTTACATACTGTTTGGTCGCCGGTGGTACTGGTCCAATGGACGAATACTTAATTGCTGGTTTGGCTGAAATCA

ACTCCACTTTCGACTTGTCTCCATCTTGGTACGTTGAAGCTTTGAAGTACATCAAAGCCAACCACGGTTTATCAGGTCAAGCTGCTGTCGAAGCTAACGCTTACATTGACTACGCTATC

AATGCTCTATCCTGA

CpcA(new)

Usage and Biology

CpcA gene encodes α-phycocyanin, a light-harvesting photosynthetic bile pigment-protein from the phycobiliprotein complex (phycobilisome, PBS). Phycocyanin is the major phycobiliprotein in the PBS rod.

Optimized

we mutate the cpc A to find the key amino acid of increasing the heat stability.Here are the mutation steps: First of all, we compared the amino acid sequence of ordinary phycocyanin with that of heat-resistant phycocyanin to find out the difference between them in the sequence. [Aa that needs mutation is marked in blue (40 in total):]

5	10	15	20	25	30	35	40
MKTPI	TEAIA	AADTQ	GRFLS	NTELQ	AADGR	FKRAV	ASMEA
ARALT	NNAQS	LIDGA	AQAVY	QKFPY	TTTMQ	GSQYA	STPEG
KAKCA	RDIGY	YLRMV	TYCLV	AGGTG	PMDEY	LIAGL	AEINS
TFDLS	PSWYV	EALKY	IKANH	GLSGQ	AAVEA	NAYID	YAINA	LS

And then used I-Mutant website for operation. The following are the operation steps on the website:
1.

Protein sequence-----Enter
2.

[ Protein sequence ]:

MKTPITEAIAAADTQGRFLSNTELQAADGRFKRAVASMEAARALTNNAQSLIDGAAQAVYQKFPYTTTMQGSQYASTPEGKAKCARDIGYYLRMVTYCLVAGGTGPMDEYLIAGLAEINSTFDLSPSWYVEALKYIKANHGLSGQAAVEANAYIDYAINALS

[ Position ]: (For example, if the first AA to be mutated is "I", fill in "5")
[ New Residue ]: Fill in the original amino acid to be mutated
[ Temperature ]: 25
[ Ph ]: 7
[ Prediction ]: (DDG)
[ E-mail ]: (Fill in your email, the result will be sent to the email)

Negative DDG means that mutation improves stability. Three New Residue records with the largest absolute value of negative values are selected from 19 mutations, such as 5 I--H, T and Y (" 5 "is the position of mutation AA in the whole sequence, I is the original AA, and the absolute value of negative values of H, T and G decreases successively). Here are our results.

5 I--D-2.1 G-2.01 W-1.74
9 I--G-2.66 P-2.36 D-2.32
10 A--P-2.15 K-1.59 Y-1.56
11 A--P-1.80 K-1.53 Y-1.31
14 T--P-1.62  G-0.93  A-0.86
21 N--G-1.93  K-1.46  A-1.35
26 A--P-2.15  G-2.10  K-1.96
28 D--G-2.11  P-1.76  K-1.69
31 F--G -3.53 D-2.27 A -2.09
32 K--D -1.78  G-1.15 T-0.99
33 R--S-1.46 K -1.37   D -1.32
  35 V--G-2.75  F -1.80 D -1.80
37  S--G-1.28  P-0.71   D-0.60
39  E--G-1.66   K-0.94 D-0.51
38  M--G-3.40   K-2.23 A-1.45
42  R--S-1.83   D-1.38 K-1.30
46 N--G-2.23 D-1.38 H-1.30
52 I--G-3.63 T-3.61 S-2.93
53 D--P-1.12 G-0.62 A-0.41
61 Q--P-1.33 S-1.24 N-1.12
68 T--G-2.22 P-1.11 S-0.83
69 M--G-1.79 S-0.83 K-0.53
72 S--G-1.32 C-0.60 P-0.14
73 Q--N-1.33 G-1.1 S-0.66
74 Y--G-3.83 A-2.13 S-1.57
76 S--G-0.97 P-0.09
77 T--G-2.08 P-1.11 Q-0.52
78 P--G-1.51 A-1.22 D-1.22
79 E--G-1.46 K-1.29 A-0.86
82 A--G-1.29 S-1.09 T-1.08
94 M--G-2.61 S-2.16 K-1.74
107 M--G-1.75 T-1.50 A-1.37
115 L--T-4.84 D-4.39 G-4.17
116 A--K-1.88 H-1.82 T-1.82
120 S--C-1.66 T-1.56 G-1.44
145 Q--N-1.25 G-1.19 P-1.18
147 A--T-1.44 P-1.32 G-1.27
148 V--G-2.62 F-1.72 P-1.66
152 A--T-1.55 P-1.40 H-1.16
154 I--T-3.69 G-3.58 D-3.14

Sequence

MKTPDTEAGPPADPQGRFLSGTELQPAGGRGDSAGAGGGAASALTGNAQSLGPGAAQAVYPKFPYTTGGQGGNGAGGGGGKGKCARDIGYYLRGVTYCLVAGGTGPGDEYLIAGTKEINSCTFDLSPSWYVEALKYIKANHGLSGNATGEANTYTDYAINALS

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]