SUI Chang-hai,WANG Lin.Cloning and Bioinformatics Analysis of Soybean Proteinase Inhibitor Gene[J].HEILONGJIANG AGRICULTURAL SCIENCES,2016,(11):1-3.[doi:10.11942/j.issn1002-2767.2016.11.00]
大豆蛋白酶抑制剂基因的克隆及生物信息学分析
- Title:
- Cloning and Bioinformatics Analysis of Soybean Proteinase Inhibitor Gene
- 文章编号:
- 11-0001-03
- Keywords:
- soybean; Jiyu 47; proteinase inhibitor
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 为了研究使用安全的抗虫蛋白，以市售吉育47种子为材料，利用RT-PCR方法，从大豆吉育47萌发种子提取RNA，以该RNA为模板进行反转录并进行PCR，扩增大豆吉育47蛋白酶抑制剂基因并进行生物信息学分析与表达载体的构建。结果表明：克隆到长252 bp的完整编码基因序列，编码84个氨基酸残基；经测序与同源比对鉴定该基因与已报道基因相似性高度一致；以pCAMBIA3301为基础构建了植物表达载体pCAMBIA3301PI并转化入农杆菌EHA105中。
- Abstract:
- In order to study the safety of insecticidal protein,with commercially available Jiyu 47 seeds as materials,total RNA was extracted,reverse transcription was carried out with the RNA as template,and amplification Jiyu 47 protease inhibitor gene by RT-PCR,the end bioinformatics analysis and construction of expression vector were carried out. The results showed that 252 bp complete sequences was cloned,it encoded 84 amino acids; sequencing and Blast with reported gene highly similar; expression vector pCAMBIA3301PI was constructed by pCAMBIA3301 and transformed into Agrobacterium EHA105.
参考文献/References:
[1]El-Hady E A A,Habiba R A. Effect of soaking and extrusion conditions on antinutrients and protein digestibility of legume seeds[J]. Lebensmittel-Wissenschaft und-Technologie,2003,36(3):285-293.
[2]Egounlety M,Aworh O C. Effect of soaking,dehulling,cooking and fermentation with Rhizopus oligosporus,on the oligosaccharides,trypsin inhibitor,phytic acid and tannins of soybean (Glycine max Merr.),cowpea (Vigna unguiculata L.Walp) and groundbean(Macrotyloma geocar)[J].Journal of Food Engineering,2003,56(2-3):249-254. [3]Lipardi C,Wei Q,Paterson B M. RNAi as random degradative PCR:siRNA primers convert mRNA into dsRNAs that are degraded to generate new siRNAs[J]. Cell,2001,107(3):297-307. [4]Matzke M,Matzke A J M,Kooter J M. RNA: Guiding gene silencing[J]. Science,2001,293(5532):1080-1083. [5]Nyk nen A,Haley B,Zamore P D. A requirements and small interfering RNA structure in the RNA interference pathway[J]. Cell,2001,107(3):309-321. [6]Stewart O J,Raghavan G S V,Orsat V,et al. The effect of drying on unsaturated fatty acids and trypsin inhibitor activity in soybean[J]. Process Biochemistry,2003,39(4):483-489. [7]陈吉刚,郑肖娟,龚辉,等.RNA干扰研究进展[J]. 中国预防兽医学报，2004(2):155-158. [8]丁安林,孙君明. 无胰蛋白酶抑制剂的优质大豆新品种中豆28[J]. 作物杂志，1999(3):29-29. [9]金蓓,田少君. 大豆胰蛋白酶抑制剂研究概况[J]. 粮食与油脂，2005(6):3-6. [10]严晴燕,曹凯鸣,黄伟达,等.大豆胰蛋白酶抑制剂SBTi-A2新类型Tid突变位点的初步研究[J]. 复旦学报：自然科学版， 1998(2):229-232. [11]严晴燕,曹凯鸣,徐隽,等.大豆（G．max）胰蛋白酶抑制剂SBTi－A2新类型Tix的纯化及其性质研究[J]. 复旦学报：自然科学版，1996(2):150-156. [12]杨丽杰,吴非,霍贵成,等.豆制品中胰蛋白酶抑制剂的研究[J].粮油加工与食品机械，2004(3):47-47.
相似文献/References:
[1]吕静,韩莉梅,张所有.超氧化物歧化酶模拟物（SODM）对大豆保护酶系统的影响[J].黑龙江农业科学,2013,(01):23.
LYUJing,HANLi-mei,ZHANGSuo-you.EffectsofSuperoxideDismutaseMimics(SODM)onSoybeanProtectiveEnzymeSystem[J].HEILONGJIANG AGRICULTURAL SCIENCES,2013,(11):23.
[2]吴俊彦,于晓光.黑河地区灾后补种大豆品种筛选试验[J].黑龙江农业科学,2014,(07):13.
WU Jun yan,YU Xiao guang.Soybean Varieties Screening Test for Reseeding in Heihe Region after Disaster[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):13.
[3]安咏梅,王家军,李进荣,等.大豆抗胞囊线虫的分子标记研究[J].黑龙江农业科学,2014,(07):15.
AN Yong mei,WANG Jia jun,LI Jin rong,et al.Molecular Marker Assisted Selection on Soybean Resistant to Soybean Cyst Nematode[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):15.
[4]刘兴龙,王克勤,朱立新,等.大豆蚜对大豆阶段性危害研究[J].黑龙江农业科学,2014,(06):57.
LIU Xinglong,WANG Keqin,ZHU Lixin,et al.Study on Effect of Soybean Aphid on Soybean Growth at Different Stages[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):57.
[5]赵 寅.富裕县大豆生产现状及发展建议[J].黑龙江农业科学,2014,(05):150.
ZHAO Yin.Production Situation and Suggestions of Soybean in Fuyu County[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):150.
[6]郑 旭,赵秀梅,王连霞,等.25%阿维〖DK1〗·福〖DK1〗·莠锈悬浮种衣剂病害防效试验[J].黑龙江农业科学,2014,(05):54.
ZHENG Xu,ZHAO Xiu-mei,WANG Lian-xia,et al.Field Efficacy Trial of 25% Abamectin·Thiram·Carboxin FSC Against Soybean Disease[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):54.
[7]袁明. 齐齐哈尔大豆综合试验站现代农业产业示范基地建设成效的探讨[J].黑龙江农业科学,2013,(01):143.
YUANMing.DiscussiononConstructionEffectofQiqiharSoybeanComprehensiveExperimentalStationofModernAgriculturalIndustrializationDemonstrationBase[J].HEILONGJIANG AGRICULTURAL SCIENCES,2013,(11):143.
[8]李亮,李泽宇,王丽娜,等.不同大豆品种耐盐性表现分析[J].黑龙江农业科学,2013,(02):17.
LI Liang,LI Ze-yu,WANG Li-na,et al.Performance Analysis on Salt Tolerance for Different Soybean Varieties[J].HEILONGJIANG AGRICULTURAL SCIENCES,2013,(11):17.
[9]郑新利,张丙双,寇贺.东北地区大豆生产主要问题及发展对策[J].黑龙江农业科学,2012,(02):146.
ZHENG Xin-li,ZHANG Bing-shuang,KOU He.Main Problems and Developing Countermeasuresof Soybean in Northeast China[J].HEILONGJIANG AGRICULTURAL SCIENCES,2012,(11):146.
[10]孙羽,冯延江,王麒,等.玉米茬原垄卡种大豆对其生长及产量的影响[J].黑龙江农业科学,2014,(03):26.
SUN Yu,FENG Yan jiang,WANG Qi,et al.Effect of Original Ridge Tillage on the Growth and Yield of Soybean[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(11):26.
备注/Memo
收稿日期：2016-09-23