CAJ | 학술논문

通过分子生物学方法,克隆得到锯缘青蟹（Scylla serrata）精氨酸激酶（Arginine Kinase,AK）开放阅读框基因序列.测序结果显示：该开放阅读框基因的序列长度为1 071 bp,编码356个氨基酸残基;该序列登录Genbank（GQ：851626）,序列比对结果显示,锯缘青蟹精氨酸激酶与凡纳滨对虾（Litopenaeus vannamei）精氨酸激酶的氨基酸序列同源性高达94%,与岸蟹（Carcinus maenas）、中华绒鳌蟹（Eriocheir sinensis）等甲壳类动物的精氨酸激酶也有较高的同源性,均高于90%;将克隆得到的锯缘青蟹精氨酸激酶基因与pGEX-4T-3载体连接,转化大肠杆菌BL21,经IPTG诱导得到分子质量为65 ku的融合表达蛋白（GST-AK）,经兔抗锯缘青蟹精氨酸激酶多克隆抗体的免疫印迹分析证实,GST-AK具有抗原性.
통과분자생물학방법,극륭득도거연청해（Scylla serrata）정안산격매（Arginine Kinase,AK）개방열독광기인서렬.측서결과현시：해개방열독광기인적서렬장도위1 071 bp,편마356개안기산잔기;해서렬등록Genbank（GQ：851626）,서렬비대결과현시,거연청해정안산격매여범납빈대하（Litopenaeus vannamei）정안산격매적안기산서렬동원성고체94%,여안해（Carcinus maenas）、중화융오해（Eriocheir sinensis）등갑각류동물적정안산격매야유교고적동원성,균고우90%;장극륭득도적거연청해정안산격매기인여pGEX-4T-3재체련접,전화대장간균BL21,경IPTG유도득도분자질량위65 ku적융합표체단백（GST-AK）,경토항거연청해정안산격매다극륭항체적면역인적분석증실,GST-AK구유항원성.
The open reading frame gene sequences of Mud crab arginine kinase was cloned through the molecular biology technology method.The cloning of arginine kinase gene amplified with sequence analysis showed that the cloned DNA fragments had open reading frames of 1 071 bp,and predicted to encode proteins with 356 amino acid residues.The obtained gene was deposited in Genbank under accession number of GQ 851626.Sequence alignment revealed that the Mud crab arginine kinase shared 94 % of homology to Litopenaeus vannamei,and more than 90 % homology to other shellfishes,such as Carcinus maenas,Eriocheir sinensis.Mud crab arginine kinase gene was further recombined with the vector of pGEX-4T-3 and expressed in Escherichia coli BL21.The expressed protein revealed a band of about 65 ku on SDS-PAGE.Western blot analysis using anti-Mud crab AK polyclonal antibody revealed positive reaction to the GST-AK fusion protein.