[發明專利]大腸桿菌耐熱性腸毒素STI突變體(Cys→Gly)分泌型表達載體

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• 申請號: 200910117233.9
• 申請日: 2009-02-24
• 公開（公告）號: CN101550424A
• 公開（公告）日: 2009-10-07
• 發明（設計）人: 王玉炯;李敏;趙輝;徐廣賢
• 申請（專利權）人: 寧夏大學
• 主分類號: C12N15/70
• 分類號: C12N15/70;C12R1/19
• 代理公司: 寧夏專利服務中心
• 代理人: 葉學軍

1、一種大腸桿菌耐熱性腸毒素ST₁突變體(Cys→Gly)分泌型表達載體，是以產腸毒素性大腸桿菌強毒株C83922所提取質粒為模板，利用引物分別進行重組PCR，擴增得到三種ST₁突變體--ST₁C10/G、ST₁C14/G和ST₁C17/G，繼而將其克隆至pET-22b中，獲得了相應的分泌型表達載體pESG10、pESG14、pESG17與相應的重組菌株BL21(DE3)(pESG10)、BL21(DE3)(pESG14)、BL21(DE3)(pESG17)。分泌型表達載體中，突變體基因核苷酸序列分別為：

突變體基因ST₁C10/G核苷酸序列：

GCT?CAG?GAT?GCT?AAA?CCA?GTA?GAG?TCT?TCA?AAA?GAA?AAA?ATC?ACA?CTA?GAA?TCA?AAA?AAA??60

A??Q???N??A??K??P????N???E???U???S???K????N????K???I???T???L???N???S????K????K???20

TGT?AAC?ATT?GCA?AAA?AAA?AGT?AAT?AAA?AGT?GGT?CCT?GAA?AGC?ATG?AAT?AGT?AGC?AAT?TAC?120

C??N???I??A??K??K????S???N???K????S???G???P???N????S???M???N???S????S????N???Y???40

TGC?TGT?GAA?TTG?TGT?GGT?AAT?CCT?GCT?TGT?ACC?GGG?TGC?TAT?????????????????????????180

C???C???E???L???C???G???N???P???A???C???T???G???C???Y????????????????????????????60

突變體基因ST₁C14/G核苷酸序列：

GCT?CAG?GAT?GCT?AAA?CCA?GTA?GAG?TCT?TCA?AAA?GAA?AAA?ATC?ACA?CTA?GAA?TCA?AAA?AAA??60

A??Q???N??A??K??P????N???E???U????S???K???N????K???I???T???L????N???S????K???K???20

TGT?AAC?ATT?GCA?AAA?AAA?AGT?AAT?AAA?AGT?GGT?CCT?GAA?AGC?ATG?AAT?AGT?AGC?AAT?TAC?120

C??N???I??A??K??K????S???N???K???S????G???P????N???S???M???N????S???S????N???Y???40

TGC?TGT?GAA?TTG?TGT?TGT?AAT?CCT?GCT?GGT?ACC?GGG?TGC?TAT?????????????????????????180

C???C???E???L????C???C???N???P???A???G???T???G???C???Y???????????????????????????60

突變體基因ST₁C17/G核苷酸序列：

GCT?CAG?GAT?GCT?AAA?CCA?GTA?GAG?TCT?TCA?AAA?GAA?AAA?ATC?ACA?CTA?GAA?TCA?AAA?AAA??60

A??Q???N??A??K???P???N???E???U????S???K????N???K???I???T????L???N???S???K????K???20

TGT?AAC?ATT?GCA?AAA?AAA?AGT?AAT?AAA?AGT?GGT?CCT?GAA?AGC?ATG?AAT?AGT?AGC?AAT?TAC?120

C??N???I??A??K???K???S???N???K????S???G????P???N???S???M????N???S???S???N????Y???40

TGC?TGT?GAA?TTG?TGT?TGT?AAT?CCT?GCT?TGT?ACC?GGG?GGC?TAT?????????????????????????180

C???C???E???L????C???C???N???P???A???C???T???G???G???Y???????????????????????????60