Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (5): 306-313.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0962
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CHEN Xiao-meng(), ZHANG Xue-jing, ZHANG Huan, ZHANG Bao-jiang, SU Yan()
Received:
2022-08-04
Online:
2023-05-26
Published:
2023-06-08
Contact:
SU Yan
E-mail:1292236360@qq.com;2006au@qq.com
CHEN Xiao-meng, ZHANG Xue-jing, ZHANG Huan, ZHANG Bao-jiang, SU Yan. Prokaryotic Expression of Recombinant Bovine Mastitis Staphylococcus aureus GapC Protein and Identification of Its B-cell Epitopes[J]. Biotechnology Bulletin, 2023, 39(5): 306-313.
Fig. 1 PCR amplification of GapC gene and double digestion of recombinant plasmid A:PCR amplification of GapC gene;M1:DL 2000 DNA Marker;1-4: amplification of GapC gene,5: negative control. B:Double digestion of recombinant plasmid of GapC gene; M2:DL 5000 DNA Marker;1-4: double digestion of pET-28a-GapC
Fig. 2 Induction and purified result of recombinant GapC M: Protein marker; 1:uninduced by GapC; 2: after induced by GapC; 3: supernatant; 4: precipitation; 5: penetration; 6-7: elution
二级结构 Secondary structure | 占总氨基酸比例 Percentage in total amino acids/% |
---|---|
Hh: α-螺旋 | 31.07 |
Ee: β-片层 | 25.74 |
Tt: β-转角 | 9.47 |
Cc: 无规则卷曲 | 33.73 |
Table 1 Secondary structure prediction of recombinant GapC protein
二级结构 Secondary structure | 占总氨基酸比例 Percentage in total amino acids/% |
---|---|
Hh: α-螺旋 | 31.07 |
Ee: β-片层 | 25.74 |
Tt: β-转角 | 9.47 |
Cc: 无规则卷曲 | 33.73 |
多肽 Polypeptide | 多肽位置 Polypeptide position | 多肽序列 Polypeptide sequence |
---|---|---|
PL1 | 66-90 | RVNGKEVKSFSEPDASKLPWKDLNI |
PL2 | 95-115 | ECTGFYTDKDKAQAHIEAGAK |
PL4 | 181-208 | TGDQNTQDAPHRKGDKRRARAA AENIIP |
PL5 | 221-236 | IPEIDGKLDGGAQRVP |
PL6 | 247-268 | VVLEKQDVTVEQVNEAMKNASN |
PL7 | 264-286 | KNASNESFGYTEDEIVSSDVVGM |
PC8 | 297-308 | TRVMSVGDRQLV |
Table 2 Prediction of antigen epitope for recombinant GapC protein
多肽 Polypeptide | 多肽位置 Polypeptide position | 多肽序列 Polypeptide sequence |
---|---|---|
PL1 | 66-90 | RVNGKEVKSFSEPDASKLPWKDLNI |
PL2 | 95-115 | ECTGFYTDKDKAQAHIEAGAK |
PL4 | 181-208 | TGDQNTQDAPHRKGDKRRARAA AENIIP |
PL5 | 221-236 | IPEIDGKLDGGAQRVP |
PL6 | 247-268 | VVLEKQDVTVEQVNEAMKNASN |
PL7 | 264-286 | KNASNESFGYTEDEIVSSDVVGM |
PC8 | 297-308 | TRVMSVGDRQLV |
Fig. 7 Immunized rabbit serum antibody levels and titer detection results of recombinant GapC A:Immunized rabbit serum antibody levels detection results. B:Immunized rabbit serum antibody levels detection results, and different letters indicate significant differences between groups(P<0.05), the same below
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