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用pGEMT-easy质粒载体构建了经白粉茵诱导0.5h、1.0h、12h、24h和48h的小麦-黑麦1BL/1RS易位系99/2439叶片cDNA文库1个.文库宿主茵为E. coliDH10B,诱导文库平均插入片段为1kb左右. 相似文献
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Qin Lu Jian-ping Lu Xiao-dong Li Xiao-hong Liu Hang Min Fu-cheng Lin 《浙江大学学报(自然科学英文版)》2008,9(7):511-519
In this study the MTP1 gene, encoding a type III integral transmembrane protein, was isolated from the rice blast fungus Magnaporthe oryzae. The Mtp 1 protein is 520 amino acids long and is comparable to the Ytp 1 protein of Saccharomyces cerevisiae with 46% sequence similarity. Prediction programs and MTP1-GFP (green fluorescent protein) fusion expression results indicate that Mtp 1 is a protein located at several membranes in the cytoplasm. The functions of the MTP1 gene in the growth and development of the fungus were studied using an MTP1 gene knockout mutant. The MTP1 gene was primarily expressed at the hyphal and conidial stages and is necessary for conidiation and conidial germination, but is not required for pathogenicity. The Amtpl mutant grew more efficiently than the wild type strain on non-fermentable carbon sources, implying that the MTP1 gene has a unique role in respiratory growth and carbon source use. 相似文献
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利用马铃薯卷叶病毒(PLRV)复制酶基因3′端长约600bp的cDNA,用缺口平移方法进行32P同位素标记,制备成cDNA探针,通过核酸斑点杂交,对提纯的马铃薯卷叶病毒(PLRV)、病毒RNA、PLRV感染的马铃薯汁液,表达PLRVCP基因的马铃薯叶片及块茎芽进行了检测.结果表明,32P标记的复制酶基因cDNA探针特异性强、灵敏度高.可测得提纯病毒的最低含量为408ng/ml,病毒RNA最低量为27.8pg/ml,未转CP基因接种PLRV的马铃薯叶片提取液的最高稀释度为1375.接种PLRV的转CP基因的抗性马铃薯块茎芽和叶片提取液的最高稀释度为0~1/15.此探针和只转入病毒外壳蛋白基因,不接种PLRV的转基因马铃薯汁液不发生反应.表明,探针只与PLRV基因组RNA特异反应,而不与外壳蛋白基因及其mRNA反应,从而为表达CP基因的转基因马铃薯中PLRV的检测和抗病性鉴定建立了一种可靠的灵敏的分子生物学技术.此项技术已用于转CP基因马铃薯Desire,Favorita,乌盟601和虎头的抗病性鉴定 相似文献
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Weidong Yong Kang Chong Tiebing Liang Zhihong Xu Kehui Tan Zhiqing Zhu 《科学通报(英文版)》1999,44(14):1289-1294
Based on the cDNA fragment sequence of vernalization-related geneverc203 cloned by differential screening in our lab, the 5′ primer has been designed. The cDNA 3′ end ofver203 gene (1 197 bp) has been cloned by the RACE method. And it is identified by Northern blotting that its expression is special
in vernalization treatment. After comparing the sequence in the nucleotide sequence databases of Genbank, EMBL and DDBJ, the
gene has homology withHordeum vulgare jesmonate-induced protein gene. It is suggested that this gene might be related to the signal transduction mediated by jamonate. 相似文献
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A new method designated cDNA array was developed by hybridization of quantitatively arrayed DNA samples isolated randomly
from a cDNA library with probes reverse-transcribed from mRNAs of different sources or treatments. The gene expression patterns
of 1 000 randomly chosen clones from an Arabidopsis library were analyzed with green seedlings versus suspension cells and
seedlings irradiated under UV light. Northern blot and sequence analysis of some differentially expressed clones confirmed
the results revealed by cDNA array, indicating that this method is efficient and reliable to monitor gene expression. 相似文献