中国比较医学杂志
中國比較醫學雜誌
중국비교의학잡지
CHINESE JOURNAL OF COMPARATIVE MEDICINE
2014年
11期
33-40
,共8页
许琴%董翔%李建瑛%许永华%任秀梅%赵彦斌%白杰英%孙兆增%曾林%胡仲明
許琴%董翔%李建瑛%許永華%任秀梅%趙彥斌%白傑英%孫兆增%曾林%鬍仲明
허금%동상%리건영%허영화%임수매%조언빈%백걸영%손조증%증림%호중명
比格犬%雌激素受体β%剪接异构体
比格犬%雌激素受體β%剪接異構體
비격견%자격소수체β%전접이구체
Beagles%Estrogen receptorβ%Splicing isoforms
目的：在比格犬下丘脑-垂体-性腺轴上筛查雌激素受体β的剪接异构体。方法针对ERβmRNA CDS序列的八个外显子，两两外显子依次组合设计引物，运用PCR法，以比格犬下丘脑、垂体、卵巢、子宫组织cDNA为模板扩增相应序列，测序后在NCBI网站比对，确定是目的基因后，用DNAMAN软件比对分析及手工校对，获得ERβ的剪接异构体。结果获得到了比格犬ERβ的四个剪接异构体，分别是第4外显子完整缺失的ERβ剪接异构体（缺失300 bp），部分第4与部分第5外显子组合缺失的两个组合型缺失的ERβ剪接异构体（各缺失334 bp，265 bp），以及第七外显子完整缺失的ERβ剪接异构体（缺失181 bp）。第4外显子完整缺失的剪接异构体和第7外显子完整缺失的剪接异构体获得了全长编码序列，另两个剪接异构体为部分编码序列。结论研究获得了比格犬ERβ的四个剪接异构体，为后续研究其在比格犬生殖调控中的作用机制打下了基础。
目的：在比格犬下丘腦-垂體-性腺軸上篩查雌激素受體β的剪接異構體。方法針對ERβmRNA CDS序列的八箇外顯子，兩兩外顯子依次組閤設計引物，運用PCR法，以比格犬下丘腦、垂體、卵巢、子宮組織cDNA為模闆擴增相應序列，測序後在NCBI網站比對，確定是目的基因後，用DNAMAN軟件比對分析及手工校對，穫得ERβ的剪接異構體。結果穫得到瞭比格犬ERβ的四箇剪接異構體，分彆是第4外顯子完整缺失的ERβ剪接異構體（缺失300 bp），部分第4與部分第5外顯子組閤缺失的兩箇組閤型缺失的ERβ剪接異構體（各缺失334 bp，265 bp），以及第七外顯子完整缺失的ERβ剪接異構體（缺失181 bp）。第4外顯子完整缺失的剪接異構體和第7外顯子完整缺失的剪接異構體穫得瞭全長編碼序列，另兩箇剪接異構體為部分編碼序列。結論研究穫得瞭比格犬ERβ的四箇剪接異構體，為後續研究其在比格犬生殖調控中的作用機製打下瞭基礎。
목적：재비격견하구뇌-수체-성선축상사사자격소수체β적전접이구체。방법침대ERβmRNA CDS서렬적팔개외현자，량량외현자의차조합설계인물，운용PCR법，이비격견하구뇌、수체、란소、자궁조직cDNA위모판확증상응서렬，측서후재NCBI망참비대，학정시목적기인후，용DNAMAN연건비대분석급수공교대，획득ERβ적전접이구체。결과획득도료비격견ERβ적사개전접이구체，분별시제4외현자완정결실적ERβ전접이구체（결실300 bp），부분제4여부분제5외현자조합결실적량개조합형결실적ERβ전접이구체（각결실334 bp，265 bp），이급제칠외현자완정결실적ERβ전접이구체（결실181 bp）。제4외현자완정결실적전접이구체화제7외현자완정결실적전접이구체획득료전장편마서렬，령량개전접이구체위부분편마서렬。결론연구획득료비격견ERβ적사개전접이구체，위후속연구기재비격견생식조공중적작용궤제타하료기출。
Objective To screen the splicing isoforms of estrogen receptor βin the Beagle hypothalamic -pituitary -gonadal axis.Methods For ERβmRNA CDS sequence of eight exons, primers were designed confined to the CDS sequences of two sequential exons.Beagle hypothalamus, pituitary, ovary and uterus tissue cDNA were used as template, and corresponding sequences were amplified by PCR.PCR products were sequenced and aligned in the NCBI web site.The correct gene was then analyzed with DNAMAN comparative analysis software and handwork checking up, thus got the ERβsplicing isoforms of Beagle. Results Four beagle ER beta splicing isomers were obtained:exon 4 complete skipping ER βisomer (300 bp missing), two kind of Beagle ERβisoforms with partial exon 4 and partial exon 5 complicated missing (isoformⅠ334 bp missing and isoformⅡ265 bp missing), and exon 7 complete missing ERβsplicing isoforms (181 bp missing).Exon 4 complete skipping and exon 7 complete missing isomers had been obtained full length coding sequence, and the other two splicing isomers were partial coding sequence.Conclusion This project gained four ERβsplicing isomers of Beagle, and that will lay an important foundation for further study of their roles in the Beagle reproductive regulation mechanism.
