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提高灿稻基因枪转化频率的研究 总被引:2,自引:0,他引:2
为了提高灿稻的基因枪转化频率,以14种重要的灿稻栽培品种为材料,对其胚性愈伤组织的诱导与再生的培养条件,转化细胞的筛选方案以及基因枪轰击参数进行了比较研究,建立起了较高优化的基因枪转化系统,并获得了一批有价值的转化植株。 相似文献
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将小鹅瘟病毒VP3基因疫苗(pcDNA-GPV-VP3)分别以每只1μg、3μg和6μg的基因枪轰击免疫30日龄四川白鹅,用免疫组织化学法检测pcDNA-GPV-VP3在鹅体内各组织器官表达情况;用间接ELISA检测免疫鹅血清中GPV抗体滴度.结果:①各剂量免疫组1d时能在免疫部位皮肤,3d时能在心脏/十二指肠/空肠/盲肠/肝脏,7d时在回肠/直肠检测到pcDNA-GPV-VP3的表达;7d表达最多;表达可持续至35~63d;表达产物的数量和表达持续时间总体规律为6μg组>3μg组>1μg组.②免疫后第7d血清抗体开始升高,21d达到最大值,免疫后第14~217d极显著(P<0.01)高于PBS和空白质粒对照;体液免疫效果存在一定的剂量依赖.研究表明,pcDNA-GPV-VP3基因枪轰击免疫雏鹅后能在免疫部位皮肤、心肌、肝脏和各肠段中表达并能够诱导雏鹅产生良好的体液免疫. 相似文献
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β-半乳糖苷酶基因(lacZ)在大型经济海藻裙带菜中的瞬间表达 总被引:3,自引:0,他引:3
用高压氦气式基因枪将SV40启动子驱动下的β-半乳糖苷酶基因(lacZ)导入大型经济海藻裙带菜不同部位的组织切块中,48小时后染色检测,在假根、孢子叶和叶片中均检测到了lacZ的瞬间表达,研究还发现,对于裙带菜组织块的基因枪法转化,压强1300psi优于1100psi。另外在裙带菜中没有发现半乳糖苷酶的染色本底。本文结果提示:基因枪法是有效的裙带菜遗传转化方法;lacZ可以作为裙带菜基因工程研究的报告基因,SV40启动子能有效驱动外源基因在裙带菜中表面,没有组织特异性。这是lacZ在裙带菜中表面的首次报道,结果可为裙带菜基因工程育种研究提供方法学基础。 相似文献
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采用PCR方法从BMP-1的cDNA中克隆了BMP-1成熟肽的编码基因,删除了BMP-1前体分子N端的信号肽序列和C端的其他序列。用HindⅢ消化1.3Kb的PCR产物,将0.84和0.46Kb两片段分别克隆到pUC载体中进DNA序列分析。分别酶切回收EcoR-HindⅢ和HindⅢ-BamHⅠ两目的基因片段,与大肠杆菌表达载体pBV-220进行退火连接,使插入基因受控于P_RP_L启动子。将含有BMP-1成熟肽编码基因重组表达质粒pBVBMP-1转化大肠杆菌宿主细胞进行温度诱导表达。结果表明,经42℃热诱导后,大肠杆菌能以包涵体形式表达BMP-1成熟肽,其分子量约为52kDa。 相似文献
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小白鼠心肌核DNA片段中的基因表达的分子开关的研究 总被引:4,自引:0,他引:4
原位染色发现,LDH(1-5)…nNAD^ (LDH:乳酸脱氢酶,NAD^ ,氢化型辅酶)可以进入核孔,可能与自己编码基因特异结合,体外表达实验中,稀释心肌DNA片段,促进LDH/DNA表达LDH(1-5),LDH(1-5)表达量与LDH/DNA中可解离的LDH(1-5)量呈正相关性。同位素^14C标记L if impglk ty LDH(1-5)…NAD^ 可以抑制LDH/DNA体外表达,LDH(1-5)表达抑制量与加入LDH(1-5)…nNAD^ 量呈正相关性,AFM直接观察到活性基因两头的调控序列可能结合自己编码的蛋白等活性因子,基因表达与调控的分子开关主要是自己编码的蛋白等活性因子,展示了未来运用AFM和同位素标记法相结合研究生物学反应的前景。 相似文献
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Relatively few studies have examined bacterial responses to the reduced gravity conditions that are experienced by bacteria
grown in space. In this study, whole genome expression of Escherichia coli K12 under clinorotation (which models some of the conditions found under reduced gravity) was analyzed. We hypothesized that
phenotypic differences at cellular and population levels under clinorotation (hereafter referred to as modeled reduced gravity)
are directly coupled to changes in gene expression. Further, we hypothesized that these responses may be due to indirect effects
of these environmental conditions on nutrient accessibility for bacteria. Overall, 430 genes were identified as significantly
different between modeled reduced gravity conditions and controls. Up-regulated genes included those involved in the starvation
response (csiD, cspD, ygaF, gabDTP, ygiG, fliY, cysK) and redirecting metabolism under starvation (ddpX, acs, actP, gdhA); responses to multiple stresses, such as acid stress (asr, yhiW), osmotic stress (yehZYW), oxidative stress (katE, btuDE); biofilm formation (lldR, lamB, yneA, fadB, ydeY); curli biosynthesis (csgDEF), and lipid biosynthesis (yfbEFG). Our results support the previously proposed hypothesis that under conditions of modeled reduced gravity, zones of nutrient
depletion develop around bacteria eliciting responses similar to entrance into stationary phase which is generally characterized
by expression of starvation inducible genes and genes associated with multiple stress responses. 相似文献
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