中国奶牛
中國奶牛
중국내우
CHINA DAIRY CATTLE
2011年
20期
1-5
,共5页
肖海霞%托乎提·阿及德%田可川%刘明军%陈从英
肖海霞%託乎提·阿及德%田可川%劉明軍%陳從英
초해하%탁호제·아급덕%전가천%류명군%진종영
RACE技术%驴%DGAT1%基因3′端%序列分析
RACE技術%驢%DGAT1%基因3′耑%序列分析
RACE기술%려%DGAT1%기인3′단%서렬분석
RACE technique%Donkey%DGAT1 gene%3′end cDNA%Sequence analysis
DGAT1(脂酰辅酶A:二脂酰甘油酰基转移酶)基因是产奶性状的一个重要功能侯选基因。由于通过预测奶牛DGAT1基因序列与马属动物DGAT1基因序列具有98%的同源性,本试验从影响牛产奶性状的DGAT1基因出发,以驴乳腺组织的RNA为模板,按照不同物种DGAT1基因的相似性设计特异引物,运用PCR和3′RACE技术扩增并获得了特异片断,特异片段回收纯化连接到pUCmT Vector载体后,转化到大肠杆菌中并筛选阳性菌落;提取质粒进行测序,结果发现该段序列与预期的目标一致,通过与其他动物同源性比较分析说明已首次克隆到驴DGAT1基因3′端。
DGAT1(脂酰輔酶A:二脂酰甘油酰基轉移酶)基因是產奶性狀的一箇重要功能侯選基因。由于通過預測奶牛DGAT1基因序列與馬屬動物DGAT1基因序列具有98%的同源性,本試驗從影響牛產奶性狀的DGAT1基因齣髮,以驢乳腺組織的RNA為模闆,按照不同物種DGAT1基因的相似性設計特異引物,運用PCR和3′RACE技術擴增併穫得瞭特異片斷,特異片段迴收純化連接到pUCmT Vector載體後,轉化到大腸桿菌中併篩選暘性菌落;提取質粒進行測序,結果髮現該段序列與預期的目標一緻,通過與其他動物同源性比較分析說明已首次剋隆到驢DGAT1基因3′耑。
DGAT1(지선보매A:이지선감유선기전이매)기인시산내성상적일개중요공능후선기인。유우통과예측내우DGAT1기인서렬여마속동물DGAT1기인서렬구유98%적동원성,본시험종영향우산내성상적DGAT1기인출발,이려유선조직적RNA위모판,안조불동물충DGAT1기인적상사성설계특이인물,운용PCR화3′RACE기술확증병획득료특이편단,특이편단회수순화련접도pUCmT Vector재체후,전화도대장간균중병사선양성균락;제취질립진행측서,결과발현해단서렬여예기적목표일치,통과여기타동물동원성비교분석설명이수차극륭도려DGAT1기인3′단。
DGAT1(acyl CoA: diacylgycerol acyltransferase) gene is the most important functional candidate gene for milk production traits.Because the bovine DGAT1 gene has 98%homology with the predicted Equus animals.In this experiment,based on bovine DGAT1 gene,we used the RNA of donkey lacteal gland tissue to make the template and designed the special primers according to different species' DGAT1 homology.The experiment obtained the amplified fragment using PCR and RACE technique.The amplified fragment was purified and inserted into pUCmT Vector then transformed into colibacillus cells.The positive recombinant plasmid was extracted and was sequenced to analyse its sequence.The result showed that this sequence was consistent with expected aim,we have firstly cloned the 3' end cDNA sequence of donkey DGAT1 gene by the homology analysis of other animals.