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Fine Mapping of the Blast Resistance Gene P/75, Linked to Pii, on Rice Chromosome 9 被引量:17
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作者 潘庆华 胡珍娣 +1 位作者 谷坂隆俊 王玲 《Acta Botanica Sinica》 CSCD 2003年第7期871-877,共7页
The gene Pi15 for resistance of rice to Magnaporthe grisea was previously identified as being linked to the gene Pii. However, there is a debate on the chromosomal position of the Pii gene, because it was originally m... The gene Pi15 for resistance of rice to Magnaporthe grisea was previously identified as being linked to the gene Pii. However, there is a debate on the chromosomal position of the Pii gene, because it was originally mapped on chromosome 6, but recent work showed it might be located on chromosome 9. To determine the chromosomal location of the Pi15 gene, a linkage analysis using molecular markers was performed in a F2 mapping population consisting of 15 resistant and 141 susceptible plants through bulked-segregant analysis (BSA) in combination with recessive-class analysis (RCA). Out of 20 microsatellite markers mapped on chromosomes 6 and 9 tested, only one marker, RM316 on chromosome 9, was found to have a linkage with the Pi15 gene with a recombination frequency of (19.1 ±3.7)%. To confirm this finding, four sequence-tagged site (STS) markers mapped on chromosome 9 were tested. The results suggested that marker G103 was linked to the Pi15 gene with a recombination frequency of (5.7±2.1)%. To find marker(s) more closely linked to the Pi15 gene, random amplified polymorphic DNA (RAPD) analysis was performed. Out of 1 000 primers tested, three RAPD markers, BAPi15486, BAPi15782 and BAPil5844 were found to tightly flank the Pi15 gene with recombination frequencies of 0.35%, 0.35% and 1.1%, respectively. These three RAPD markers should be viewed as the starting points for marker-aided gene pyramiding and cloning. A new gene cluster of rice blast resistance on chromosome 9 was also discussed. 展开更多
关键词 bulked-segregant analysis (BSA) MICROSATELLITE rice (Oryza sativa) Magnaporthe grisea RAPD recessive-class analysis (RCA) sequencetagged site (STS)
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热胁迫后家蝇幼虫cDNA文库构建与随机EST测序分析 被引量:12
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作者 刘红美 张洁 +4 位作者 王贇 付萍 国果 张勇 吴建伟 《免疫学杂志》 CAS CSCD 北大核心 2010年第9期772-775,共4页
目的构建家蝇幼虫cDNA文库,筛选重要免疫因子。方法采用热激法处理家蝇3日龄幼虫,提取处理后培养24h的家蝇幼虫总RNA,运用SMART技术构建cDNA文库并随机挑选克隆进行EST测序分析。结果 cDNA文库的滴度为1.2×106。PCR扩增鉴定显示所... 目的构建家蝇幼虫cDNA文库,筛选重要免疫因子。方法采用热激法处理家蝇3日龄幼虫,提取处理后培养24h的家蝇幼虫总RNA,运用SMART技术构建cDNA文库并随机挑选克隆进行EST测序分析。结果 cDNA文库的滴度为1.2×106。PCR扩增鉴定显示所选的30个克隆均含有重组的插入片段,大小均大于1kb。随机挑取192个克隆进行表达序列标签(EST)测序,有效序列为171条,根据EST测序结果计算文库重组率达94%。经序列拼接得到129个Unigenes。通过序列比对发现其中72个Unigenes与GenBank中的已知基因同源,并从中鉴定出热激蛋白、变态原、转铁蛋白、几丁质酶、副肌球蛋白、丝氨酸蛋白酶抑制因子等重要免疫因子。结论高质量的家蝇幼虫cDNA文库构建成功,该文库的构建将有助于家蝇热应激免疫相关功能基因分离和进一步研究。 展开更多
关键词 家蝇 热激 CDNA文库 EXPRESSED SequenceTags(ESTs)
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