CAJ | 학술논문

本实验提取隆线溞孤雌溞和两性雌溞的可溶性蛋白进行双向电泳和质谱鉴定,分析隆线溞在两种生殖状态下蛋白质组的差异变化.聚丙烯酰胺凝胶SDS-PAGE结果表明:隆线溞在两种生殖状态下存在明显的蛋白质表达差异,孤雌溞的蛋白条带在分子量约50.6 kD、36.2 kD、32.1 kD和25.7 kD处表达量较两性雌溞明显;两性雌溞的蛋白条带在分子量约87.8 kD、67.2 kD、53.6 kD和35.5kD处表达量较孤雌溞明显,其中35.5kD的蛋白条带为两性雌溞所特有.同时取两个样品的可溶性蛋白进行双向电泳,每个样品重复四次.双向电泳图谱经银染后利用软件分析可知,隆线溞孤雌溞平均可检测到约750个蛋白质点,两性雌溞平均可检测到约720个蛋白质点.同时利用软件对凝胶上的蛋白质点进行半定量分析,发现隆线溞从孤雌生殖转化为两性生殖后有18个蛋白质点呈现显著变化,其中14个点表达量明显下降,4个点表达量显著升高.实验结果具有较好的重复性.取4个表达量显著上升的蛋白质点进行质谱分析,得到两个蛋白质点(16号和17号)的测定结果.其中16号点为一类酸性脱氢酶(2I234),它在动物生长发育的各个阶段大量表达,这类蛋白质在隆线溞生殖转化过程中表达量变化尤为显著.本研究结果表明:隆线溞在孤雌生殖和两性生殖状态下存在明显的蛋白质表达差异.
본실험제취륭선소고자소화량성자소적가용성단백진행쌍향전영화질보감정,분석륭선소재량충생식상태하단백질조적차이변화.취병희선알응효SDS-PAGE결과표명:륭선소재량충생식상태하존재명현적단백질표체차이,고자소적단백조대재분자량약50.6 kD、36.2 kD、32.1 kD화25.7 kD처표체량교량성자소명현;량성자소적단백조대재분자량약87.8 kD、67.2 kD、53.6 kD화35.5kD처표체량교고자소명현,기중35.5kD적단백조대위량성자소소특유.동시취량개양품적가용성단백진행쌍향전영,매개양품중복사차.쌍향전영도보경은염후이용연건분석가지,륭선소고자소평균가검측도약750개단백질점,량성자소평균가검측도약720개단백질점.동시이용연건대응효상적단백질점진행반정량분석,발현륭선소종고자생식전화위량성생식후유18개단백질점정현현저변화,기중14개점표체량명현하강,4개점표체량현저승고.실험결과구유교호적중복성.취4개표체량현저상승적단백질점진행질보분석,득도량개단백질점(16호화17호)적측정결과.기중16호점위일류산성탈경매(2I234),타재동물생장발육적각개계단대량표체,저류단백질재륭선소생식전화과정중표체량변화우위현저.본연구결과표명:륭선소재고자생식화량성생식상태하존재명현적단백질표체차이.
There are two different reproductive modes in daphnids: parthenogenetic and sexual reproduction, but the sexual reproduction, but the mechanism of reproductive switch has not been elucidated. In the present study, water-soluble proteins were isolated from sexual females of Daphnia (Ctenodaphnia) carinata respectively, then profiled by two-dimensional polyacrylamide gel electrophoresis and identified by mass spectrometry. Firstly, sodium dodecyl sulphate polyacrylamide istribution. More than 4 protein bands at approximately 50.6 kD, 36.2 kD, 32.1 kD and 25.7 kD were dominant in the parthenogenetic ds at approximately 87.8 kD, 67.2 kD, 53.6 kD and 35.5 kD were dominant in the sexual female sample, in which the 35.5 kD band appeared as unique to the sexual female. The results of two-dimensional polyty were obtained with four repeated assays. On average, about 750 and 720 spots were visualized respectively by sliver staining in 2D gels of parthenogenetic e reproductive mode of D. Carinata had switched from parthenogenetic to sexual reproduction, 18 proteins were shown to display significant and reproducible changes. Among them, 14 proteins were down-reguher characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry ( MALDI-TOF- MS), 2 proteins (No. 16 and No. 17) 16 protein was a branched acid dehydrogenase (2I234), which is expressed at high levels at all stages of development, especially during the transition from parthenogenetic to sexual reproduction in D. Carinata.es during the change from parthenogenetic to sexual reproduction in D. Carinata.