甜瓜响应甜瓜坏死斑点病毒侵染的cDNA文库构建

为解析甜瓜坏死斑点病毒(melon necrotic spot virus,MNSV)与寄主甜瓜的互作,筛选病毒编码的蛋白与寄主的互作因子,构建了MNSV侵染的甜瓜cDNA文库。在甜瓜品种白玫子叶期接种MNSV,待植株表现典型的系统坏死斑点症状后,收集症状明显的整株甜瓜幼苗,采用SMART(switching mechanism at 5′end of RNA transcript)技术合成cDNA,利用DSN(duplex-specific nuclease)对cDNA均一化处理后,首次构建了MNSV侵染甜瓜的cDNA文库并评价了文库的质量,文库容量为4.5×106 cfu,文库滴度为3.6×10^7 cfu·mL^-1,文库克隆重组率为100%,文库插入片段大小在500~2000 bp之间,说明文库质量很好。该文库的构建为解析MNSV与寄主互作及甜瓜响应MNSV的致病机制奠定了基础。

Intensity-modulated radiotherapy for breast cancer:a systematic review

Objective:The aim of this study was to assess the long-term effect of breast conservation with breast intensitymodulated radiotherapy (IMRT) for early-stage breast cancers.Methods:Pub Med,EMBASE,Cochrane Library,Chinese Biomedical Literature Database were searched to identify relevant original published trails,and the references of eligible studies were manually screened.Randomized controlled trails reported in any language,comparing breast IMRT with conventional radiotherapy (RT) in patients with early-stage breast cancer were eligible for inclusion.Two investigators independently assessed the quality of included trials and extracted data.The RevMan 5.0 software was used for statistical analysis.Results:Four trials involving 1440 patients were included.The results showed that in the IMRT group,the occurrence of acute moist dermatitis,the edema of the mammary gland and the occurrence of hyperpigmentation were lower than those of RT group,the pooled risk ratio value and 95% confidence interval were 0.28 (0.17-0.48),0.03 (0.00-0.21) and 0.06 (0.02-0.15) respectively.The use of IMRT did not correlate with pain and quality of life,but the presence of moist desquamation did significantly correlate with pain and reduced quality of life.Conclusion:The currently available evidence showed that IMRT significantly reduces the occurrence of moist acute dermatitis anywhere in the breast and alleviates the suffering of patients and improves the quality of life of patients.Future large-scale,high-quality,double-blind trials are needed.

Construction and quality identification of T7 recombination expression cDNA library form human lung cancer

Objective:Currently,only a limited numbers of tumor markers for non small lung cancer(NSCLC) diagnosis,new biomarker,such as serum autoantibodies may improve the early detection of lung cancer.Our objective is construction human lung squamous carcinoma and adenocarcinoma T7 phage display cDNA library from the tissues of NSCLC patients.Methods:mRNA was isolated from a pool of total RNA extract from NSCLC tissues obtained from 5 adenocarcinomas and 5 squamous carcinomas,and then mRNA was reverse transcribed into double stranded cDNA.After digestion,the cDNA was inserted into T7Select 10-3 vector.The phage display cDNA library was constructed by package reaction in vitro and plate proliferation.Plaque assay and PCR were used to evaluate the library.Results:Two T7 phage display cDNA library were established.Plaque assay show the titer of lung squamas carcinoma library was 1.8 × 106 pfu,and the adenocarcinoma library was 5 × 106 pfu.The phage titer of the amplified library were 3.2 × 1010 pfu/mL and 2.5 × 1010 pfu/mL.PCR amplifica-tion of random plaque show insert ratio were 100%(24/24) in adenocarcinoma library and 95.8% in human lung squamas carcinoma library(23/24).Insert range from 300 bp to 1 500 bp.Conclusion:Two phage display cDNA library from NSCLC were constructed.

野生型工业酿酒酵母Miseq测序方法的建立

新一代测序方法在基因组学研究应用日趋广泛,已经成为工业酿酒酵母菌株基因组学研究的重要技术平台,但对工业酿酒酵母新一代测序技术方法缺乏详细报道。Miseq是小型新一代基因组测序仪,本文报道我们自建的野生型工业酿酒酵母基因组Miseq测序方法。该方法包括：制备分离a型和α型交配型的单倍体菌株、采用电泳和分光光度法方法监控基因组文库建立、优化上机文库浓度后测序。其中电泳和分光光度法方法为首创的文库质量监控简易方法,质控检测得到酿酒酵母工业菌株测序条件为：菌株的基因组文库DNA片段大小为250-850bp且主要集中在350-550bp,文库浓度范围为7-13ng/μL;上机测序文库的优化浓度为20pM。