CAJ | 학술논문

水稻幼苗经缺铁胁迫诱导分别处理1、3、5天后,用酚法和TCA/丙酮法提取叶片中的可溶性蛋白进行双向电泳分析,从而研究在缺铁条件下叶片中蛋白表达的动态变化规律.结果显示:1.不同pH IPG胶条分离蛋白的效果不同.用pH3-10的IPG胶条进行双向电泳,经考马斯亮蓝染色后,可在胶面上检测到大约450个蛋白点,其中约有89%的蛋白是酸性蛋白.如果用pH4-7的IPG胶条进行双向电泳,则可检测到大约600个蛋白点,其中有29个蛋白是上调表达,1个蛋白是下调表达,5个蛋白是诱导特异表达.2.不同方法提取的可溶性蛋白质量不同.TCA法简单易操作,似乎对于碱性蛋白的抽提效果更好,在2-DE图像上,减性端显示的蛋白点多;但此方法所得蛋白的再溶性差.酚法提取的蛋白再溶性好,所抽提的蛋白量较大,纯度较高.
수도유묘경결철협박유도분별처리1、3、5천후,용분법화TCA/병동법제취협편중적가용성단백진행쌍향전영분석,종이연구재결철조건하협편중단백표체적동태변화규률.결과현시:1.불동pH IPG효조분리단백적효과불동.용pH3-10적IPG효조진행쌍향전영,경고마사량람염색후,가재효면상검측도대약450개단백점,기중약유89%적단백시산성단백.여과용pH4-7적IPG효조진행쌍향전영,칙가검측도대약600개단백점,기중유29개단백시상조표체,1개단백시하조표체,5개단백시유도특이표체.2.불동방법제취적가용성단백질량불동.TCA법간단역조작,사호대우감성단백적추제효과경호,재2-DE도상상,감성단현시적단백점다;단차방법소득단백적재용성차.분법제취적단백재용성호,소추제적단백량교대,순도교고.
Rice seedlings were treated in Fe-deficient hydroponic solution for 1d,3d and 5 d respectively. Then soluble proteins in rice leaves were extracted with phenolic or TCA / acetone method, and protein dynamic changes were studied on the Fe-deficient stress by 2-DE analysis. The results depicted as follows:. First, using different pH IPG gel strip leaded to different separation effectiveness. Stained with coomassie brilliant blue, nearly 450 protein spots in a 2-DE gel were detected with the pH 3～10 gradient IPG strip, among which about 89% proteins were acidic ones. However, about 600 spots were tested in a 2-DE gel with the pH4～7 gradient IPG strip. In the 2-DE gel with pH4～7 IPG strip, 29 proteins were up-regulated, only 1 protein was down-regulated, 5 proteins were specially displayed. Second, the quality and quantity of the soluble proteins were different with different extraction methods. TCA technique was easy to operate and effective to distribute basic proteins on the basic apex of 2-DE gels. But the proteins extracted by TCA were difficult to resolve thoroughly. The proteins extracted by phenolic method were more quantitative and purer than that by TCA technique.