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作 者:李明[1] 潘小玲[2] 王莉莉[1] 孙艳 冯云[1] 黄宁[1] 吴琦[1] 李绚[1] 王伯瑶[1]
机构地区:[1]四川大学华西基础医学与法医学院感染免疫研究室,成都610041 [2]四川大学华西第二医院妇产科 [3]四川生物制品研究所
出 处:《四川大学学报(医学版)》2005年第3期315-317,共3页Journal of Sichuan University(Medical Sciences)
基 金:国家自然科学基金 (批准号 3 980 0 14 6);纽约中华医学基金会 (96-681)资助
摘 要:目的 对本室从人子宫颈粘液分离纯化的1个抗菌多肽HCP- 1进行分子结构特性分析。方法 分离HCP- 1多肽,测定其氮端氨基酸序列,根据其氮端氨基酸序列密码子设计简并引物,采用3′- RACE- PCR技术扩增HCP- 1编码基因,连接到T载体进行测序。用生物软件分析测序结果。结果 以简并引物结合RACE- PCR技术成功的克隆出HCP- 1全长c DNA,BL AST分析证实其序列与HMG- 17分子相同,OMIGA软件分析发现其含有一个α-螺旋结构(疏水结构区)可能是其抗菌功能的基础。结论 HCP- 1为HMG- 17,是存在于宫颈粘液抗菌的一种抗菌多肽,其中的α-螺旋结构可能是其抗菌的结构基础。Objective To identify a new antibactrial polypeptide HCP-1 isolated from human cervical mucus. Methods HCP-1 was isolated and purified from human cervical mucus, and N-terminal sequence of HCP-1 was determined. A degenerate primer was designed according to CodeHop methods, and an 'anchor-oliga-dT primer' was used for the synthesis of cDNA. Using the degenerate primer and anchor-primer, cDNAs were amplified by PCR.The PCR products were cloned, sequenced, and analyzed by biological software. Results N-terminat sequence of HCP-1 was PKRKAEGDAK. The full length of HCP-1 cDNA was isolated and of which the sequence was the same as HMG-17. OMIGA software analysis indicated that this molecule contained an α-helix region. Conclusion The new antibacterial polypeptide isolated from human cervical mucus is HMG-17. It may play a role in the human cervical mucus and the α-helix domain may be its antibacterial activity region.
关 键 词:人子宫颈粘液 抗菌多肽cDNA克隆 结构分析
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