口腔生物医学
口腔生物醫學
구강생물의학
ORAL BIOMEDICINE
2015年
2期
83-86
,共4页
李阳%龚依伊%谢亦林%王劲松%张春梅%王松灵
李暘%龔依伊%謝亦林%王勁鬆%張春梅%王鬆靈
리양%공의이%사역림%왕경송%장춘매%왕송령
牙胚%蛋白质提取%尿素%电动研磨%小型猪
牙胚%蛋白質提取%尿素%電動研磨%小型豬
아배%단백질제취%뇨소%전동연마%소형저
Tooth germ%Protein extraction%Urea%Electro-grinding%Miniature pig
目的：比较不同蛋白质提取方法在牙胚蛋白提取中的效果，寻找适合牙齿发育高通量蛋白质组学分析的蛋白质提取方法。方法：取猪胚胎第40、50、60天第三乳磨牙牙胚，利用碱性碳酸盐溶液提取牙胚初步钙化部分的蛋白，通过聚丙烯酰胺凝胶电泳比较液氮研磨法和电动研磨器研磨法提取牙胚总蛋白效果，BCA（bicinchoninic acid）蛋白浓度测定法比较 RIPA 裂解液与尿素溶液提取牙胚总蛋白含量。结果：与液氮研磨法相比，电动研磨法提取蛋白后的聚丙烯酰胺电泳图谱蛋白条带明显更深，蛋白含量高。尿素溶液对牙胚总蛋白的提取量高于 RIPA 裂解液组，蛋白电泳分离效果均可。结论：采用电动研磨器研磨牙胚，组织损失量小，提取蛋白效果好于液氮研磨；采用尿素溶液提取牙胚总蛋白效果优于 RIPA 裂解液。初步找到适合下游高通量蛋白质组学分析的牙胚蛋白提取方法。
目的：比較不同蛋白質提取方法在牙胚蛋白提取中的效果，尋找適閤牙齒髮育高通量蛋白質組學分析的蛋白質提取方法。方法：取豬胚胎第40、50、60天第三乳磨牙牙胚，利用堿性碳痠鹽溶液提取牙胚初步鈣化部分的蛋白，通過聚丙烯酰胺凝膠電泳比較液氮研磨法和電動研磨器研磨法提取牙胚總蛋白效果，BCA（bicinchoninic acid）蛋白濃度測定法比較 RIPA 裂解液與尿素溶液提取牙胚總蛋白含量。結果：與液氮研磨法相比，電動研磨法提取蛋白後的聚丙烯酰胺電泳圖譜蛋白條帶明顯更深，蛋白含量高。尿素溶液對牙胚總蛋白的提取量高于 RIPA 裂解液組，蛋白電泳分離效果均可。結論：採用電動研磨器研磨牙胚，組織損失量小，提取蛋白效果好于液氮研磨；採用尿素溶液提取牙胚總蛋白效果優于 RIPA 裂解液。初步找到適閤下遊高通量蛋白質組學分析的牙胚蛋白提取方法。
목적：비교불동단백질제취방법재아배단백제취중적효과，심조괄합아치발육고통량단백질조학분석적단백질제취방법。방법：취저배태제40、50、60천제삼유마아아배，이용감성탄산염용액제취아배초보개화부분적단백，통과취병희선알응효전영비교액담연마법화전동연마기연마법제취아배총단백효과，BCA（bicinchoninic acid）단백농도측정법비교 RIPA 렬해액여뇨소용액제취아배총단백함량。결과：여액담연마법상비，전동연마법제취단백후적취병희선알전영도보단백조대명현경심，단백함량고。뇨소용액대아배총단백적제취량고우 RIPA 렬해액조，단백전영분리효과균가。결론：채용전동연마기연마아배，조직손실량소，제취단백효과호우액담연마；채용뇨소용액제취아배총단백효과우우 RIPA 렬해액。초보조도괄합하유고통량단백질조학분석적아배단백제취방법。
Objective:To analyze different protein extraction methods from tooth germ,and find a suitable protein extraction method for high through-put proteome analysis of tooth development.Methods:The deciduous third molars were collected from miniature pig on embryonic 40,50,60 day.The matrix samples of partial calcification tooth germ were fractionated with 50 mmol /L carbonate buffer (pH 10.8).The protein extraction effect was observed by sodium dodecylsulphate-polyacrylamide gelelectrophoresis (SDS-PAGE),pulver-ized methods under liquid nitrogen into fine powder or with an electro-grinding machine were compared.Then proteins were quantified with the method of bicinchoninic acid (BCA),RIPA lysis buffer and 8 mol /L urea were compared.Results:SDS-PAGE showed that e-lectro-grinding got higher quantity of tooth germ proteins than pulverizing under liquid nitrogen.At developing stages of tooth germ in miniature pigs,the extraction quantity of tooth germ proteins raised obviously when 8 mol /L urea replaced RIPA lysis buffer.Conclu-sions:The electro-grinding method was better for tissue utilization and 8 mol /L urea solution was suitable for extracting proteins from tooth germ with higher value of protein concentrations.The successful establishment of the extraction technique lays the foundation for the analysis methods in down-stream proteomics study.
