| 重叠延伸拼接法从基因组DNA中获得Ers-MIH1基因的编码区 |
| |
| 引用本文： | 郭豫杰,鲁维飞,韩立强,杨国宇.重叠延伸拼接法从基因组DNA中获得Ers-MIH1基因的编码区[J].河北渔业,2006(10):13-15,48. |
| |
| 作者姓名： | 郭豫杰 鲁维飞 韩立强 杨国宇 |
| |
| 作者单位： | 河南农业大学牧医工程学院,河南,郑州,450002 |
| |
| 摘 要： | 蜕皮抑制激素(molt-inhibiting hormone,MIH)产生于端髓X-器官(MTXO),储存在甲壳动物眼柄的窦腺中,抑制Y-器官蜕皮素的分泌,参与调控甲壳动物的生长和发育等重要的生理活动。提取中华绒螯蟹肌肉组织中的基因组DNA,以此为模板,采用重叠延伸拼接法(gene splicing by overlap extension,SOE)将中华绒螯蟹蜕皮抑制激素1Eriocheir sinensismolt-inhibiting hormone-1(Ers-MIH1)成熟肽基因的编码区连接起来,克隆到pMD18-T载体中,随机挑取3个阳性克隆测序,测序结果与已知的Ers-MIH1基因编码区的序列相一致。从基因组DNA中克隆该基因的方法有效地解决了取材不便给研究中华绒螯蟹蜕皮抑制激素基因所带来的限制等问题。
|
| 关 键 词： | 中华绒螯蟹 基因组DNA 蜕皮抑制激素基因1 重叠延伸拼接法 |
| 收稿时间： | 2006-07-04 |
| 修稿时间： | 2006-07-04 |
Cloning of the coding region fragment of the Ers-MIH1 gene from genomic DNA by SOE method |
| |
| Guo Yujie,Lu Weifei,Han Liqiang,Yang Guoyu.Cloning of the coding region fragment of the Ers-MIH1 gene from genomic DNA by SOE method[J].Hebei Fisheries,2006(10):13-15,48. |
| |
| Authors: | Guo Yujie Lu Weifei Han Liqiang Yang Guoyu |
| |
| Institution: | College of Animal Husbandry and Veterinary Science, Henan Agricultural University, Zhengzhou 450002, China |
| |
| Abstract: | MIH is produced in the medulla terminalis of the X-organ(MTXO) and stored in the sinus gland of the eyestalk.MIH negatively regulates the synthesis of ecdysteroids in Y-organs and plays an important role in the regulation of growth and development.Using the genomic DNA extracted from muscle tissues of the mitten crab,Eriocheir sinensis as template,the two coding region fragments of the Ers-MIH1 gene were linked by SOE method.Two pairs of primers were designed based on the complete sequence of molt-inhibiting hormone-1(Ers-MIH1) gene of the mitten crab and a part of linker peptide gene sequence was added on 3′and 5′ terminals respectively.The PCR products were mixed and re-amplified and the coding region cDNA fragment was spliced by overlap extension.The product was inserted into the vector pMD18-T.The analysis of the sequence data shows that it is identical with that of the mature peptide coding cDNA fragment of ErsMIH1 gene. |
| |
| Keywords: | Eriocheir japonica sinensis genomic DNA molt-inhibiting hormone-1 gene splicing by overlap extension(SOE) |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
