cDNA cloning,sequence analysis,and recombinant expression of akitonin beta,a C-type lectin-like protein from Agkistrodon acutus

查看全文 作　　者：Xiang-[1]dongZHA;[1]JingLIU;Kang-[2]senXU

机构地区：[1]SchoolofLifeScience,UniversityofScienceandTechnology,Hefei230026;[2]NationalInstituteforControlofPharmaceuticalsandBiologicalProducts,Beijing100050,China

出　　处：《Acta Pharmacologica Sinica》索引2004年第25卷第3期,共6页

摘　　要：AIM: To clone the cDNA of a new member of snake venom C-type lectin-like proteins, to study its structure- function relationships and to achieve its recombinant production. METHODS: PCR primers were designed based on the homology and cDNA was amplified by RT-PCR using total RNA from snake venom gland as the template. The PCR products were cloned into the plasmid pGEM-T and sequenced. The deduced protein sequence was analyzed with some bioinformatic programs. A recombinant expression plasmid was constructed using pBAD- TOPO as vector and transformed into E.coli TOP10 competent cells. RESULTS: A novel cDNA sequence encod- ing akitonin β was found and accepted by GenBank (accession number AF387100). Akitonin β consists of a typical carbohydrate recognition domain (CRD) of C-type lectins, and it is homologous with other snake venom C-type lectin-like proteins. It was predicted to be a platelet antagonist. Upon induction with arabinose rAkitonin β express- ing in E coli was achieved at a high level (superior to 150 mg/L). The recombinant fusion protein exhibited inhibitory activities on rat platelet aggregation in vitro. CONCLUSION: A new member of snake venom C-type lectin-like proteins was discovered and characterized, and an efficient recombinant expression system was estab- lished for its production. 关键词：akitonin-β CDNA 克隆 DNA序列分析 C型血凝素样蛋白急尖形蝮蛇 RT-PCR

相关文献

参考文献（29）
引证文献（4）
耦合文献（71）

参考文献(29)

引证文献(4)

耦合文献(71)

互联网出版许可证新出网证(渝)字10号全国400电话 - 免长途话费

cDNA cloning,sequence analysis,and recombinant expression of akitonin beta,a C-type lectin-like protein from Agkistrodon acutus

参考文献(29)

引证文献(4)

耦合文献(71)

用户提示