质粒类型: | 大肠杆菌表达载体 |
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启动子: | Trc |
载体标签: | N-CKS,C-His |
酶识别位点: | P3C |
载体抗性: | Ampicillin |
5' 测序引物及序列: | 5 ATC CAT GTT GCT GTT GCT CAG G 3 |
3' 测序引物及序列: | 5 GAT TTA ATC TGT ATC AGG 3 |
pTrc-CKS表达载体采用E.coliTrc启动子和蛋白质工程改造过CKS (CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase or CMP-KDO synthetase)基因融合表达,同时选用人鼻病毒14亚型3C蛋白酶专一性切割位点。CKS融合表达是美国Abbot公司的专利,CKS融合蛋白研制成功HIV和HCV重组抗原并取得FDA认证在美国上市。CKS是一个非常稳定且高度可溶的理想融合伴侣。本发明除去掉CKS C端8个氨基酸外,还将C端Met突变成Thr
CKSmut(E.coli CTP:CMP-3-deoxy-D-manno-octulosonate cytidylyltransferase or CMP-KDO synthetase)编码氨基酸序列如下(含多克隆位点):
MSFVVIIPARYASTRLPGKPLVDINGKPMIVHVLERARESGAERIIVATDHEDVARAVEAAGGEVCMTRADHQSGTE
RLAEVVEKCAFSDDTVIVNVQGDEPMIPATIIRQVADNLAQRQVGMATLAVPIHNAEEAFNPNAVKVVLDAEGYALYFSR
ATIPWDRDRFAEGLETVGDNFLRHLGIYGYRAGFIRRYVNWQPSPLEHIETLEQLRVLWYGEKIHVAVAQEVPGTGVDTP
EDGSMPDSLEVLFQGPAGSAEFSASHHHHHH
268 Amino Acids Molecular Weight 29KD