华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2013年
4期
75-78
,共4页
秦彤%王皓宇%郝海生%王栋%杜卫华%朱化彬
秦彤%王皓宇%郝海生%王棟%杜衛華%硃化彬
진동%왕호우%학해생%왕동%두위화%주화빈
围产奶牛%乳腺%内参基因%实时荧光定量PCR%geNorm程序
圍產奶牛%乳腺%內參基因%實時熒光定量PCR%geNorm程序
위산내우%유선%내삼기인%실시형광정량PCR%geNorm정서
Periparturient cows%Mammary gland%Reference gene%qRT-PCR%geNorm software
选择合适的内参基因是利用实时荧光定量PCR准确分析基因相对表达量的先决条件。选择了奶牛乳腺研究中广泛使用的7个候选内参基因,包括GAPDH、ACTB、18S rRNA、RPS9、RPS15、UXT、MYC。奶牛以精粗比不同的2种日粮饲喂,并在产后16 d采集乳腺组织。利用qRT-PCR技术探讨了候选内参基因在围产奶牛乳腺组织中的表达情况,并利用geNorm程序对数据进行分析。结果发现,UXT和RPS9表达稳定,不受日粮精粗比的影响,因此,可作为围产期奶牛乳腺组织基因表达分析中相对合适的内参基因。
選擇閤適的內參基因是利用實時熒光定量PCR準確分析基因相對錶達量的先決條件。選擇瞭奶牛乳腺研究中廣汎使用的7箇候選內參基因,包括GAPDH、ACTB、18S rRNA、RPS9、RPS15、UXT、MYC。奶牛以精粗比不同的2種日糧飼餵,併在產後16 d採集乳腺組織。利用qRT-PCR技術探討瞭候選內參基因在圍產奶牛乳腺組織中的錶達情況,併利用geNorm程序對數據進行分析。結果髮現,UXT和RPS9錶達穩定,不受日糧精粗比的影響,因此,可作為圍產期奶牛乳腺組織基因錶達分析中相對閤適的內參基因。
선택합괄적내삼기인시이용실시형광정량PCR준학분석기인상대표체량적선결조건。선택료내우유선연구중엄범사용적7개후선내삼기인,포괄GAPDH、ACTB、18S rRNA、RPS9、RPS15、UXT、MYC。내우이정조비불동적2충일량사위,병재산후16 d채집유선조직。이용qRT-PCR기술탐토료후선내삼기인재위산내우유선조직중적표체정황,병이용geNorm정서대수거진행분석。결과발현,UXT화RPS9표체은정,불수일량정조비적영향,인차,가작위위산기내우유선조직기인표체분석중상대합괄적내삼기인。
Selection of appropriate reference genes for normalization of quantitative PCR data for genes of inter -est is critical for interpretation of results .Seven candidate reference genes including GAPDH, ACTB,18S rRNA, RPS9,RPS15,UXT and MYC were selected in this research.Cows were fed different forage to concentrate (F:C)ra-tio dietary and mammary biopsies were taken at 16 day postpartum .The mRNA expression levels of seven candidate reference genes in mammary tissue were detected by qRT-PCR.Gene copy number was determined using absolute quantification ,and then the expression stability of these genes were analysed by geNorm software .These results sug-gested that UXT and RPS9 showed less variability between the cows and were unaffected by dietary F:C ratio and, therefore ,could be used to normalize mRNA levels of mammary in periparturient cows .