S1核酸酶突变检测法的优化

优化了S1核酸酶突变检测体系,结果表明(1)扩增法比杂交法产生异源双链DNA更节省时间;(2)PCR产物可以直接作为突变检测的底物;(3)适当降低反应温度、适当增加反应缓冲液中的NaCl浓度、适当缩短反应时间可提高突变检测效果。     

中国西门塔尔牛第6号染色体上部分微卫星的特征

利用微卫星技术对牛第6号染色体上9个微卫星位点进行了研究,检测其在中国西门塔尔牛育种核心群中152头母牛的多态性。本试验9个位点中,BM4528为单态,BM4203属于中度多态位点(0.5>PIC>0.25),而其它的7个位点均是高度多态位点(PIC>0.5)。     

家畜繁殖生物技术研究现状及趋势

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。     

鹅肥肝形成的分子机理研究进展

鹅肥肝同鲟鱼翅酱、黑菌(块菰)被西方人誉为世界三大美食珍品。本文综述了近来年鹅肥肝形成的生化机理。不同鹅种产肝性能差异的原因，可能在生化及分子标记方面的研究进展。     

从甜瓜子叶中提取总DNA

用分子标记方法检测甜瓜杂交种子纯度,如果从真叶中提取DNA,检测周期至少需要15d。为了缩短检测周期,我们分别以甜瓜的成熟干种子、吸胀水后“露白”种子、3d龄刚转绿子叶、6d龄子叶、9d龄子叶和12d龄子叶为材料,进行了提取总DNA的研究。结果表明:除了干种子和吸胀水后“露白”种子外,其他的材料都可以提取到DNA,但是DNA的质量因子叶日龄不同而存在很大的差异。不同日龄的子叶中,以3d龄子叶提取的DNA质量好。子叶6d龄时,提取的DNA质量次于3d龄,少量DNA开始出现降解,以后随着子叶日龄的增加,DNA降解加重。另外,与以真叶为材料提取的DNA样品相比较,子叶DNA样品中的蛋白质等杂质含量高,应增加氯仿抽提纯化次数。     

Role of polymerase β in DNA metabolism and genomic instability

The major role of DNA polymerase β was thought to be limited in its involvement in short patch base excision repair by removing 5’-deoxyribose phosphate and base insertion. However, the recent researches indicate that polymerase β might take part in a wide spectrum of DNA metabolism reactions, including long patch base excision repair, DNA replication, recombination, meiosis and transleisional DNA synthesis. Because of its wide and important cellular function, an inappropriate intracellular polymerase β level might be associated with genomic instability. Down-regulation or mutation of polymerase β is mutagenic due to deficient in DNA repair, while overexpression of this error-prone β polymerase might perturb the normal function of other accurate polymerases and cause genomic instability as well.     

Comparison of Resistance to Tomato Leaf Curl Virus (India) and Tomato Yellow Leaf Curl Virus (Israel) among Lycopersicon Wild Species, Breeding Lines and Hybrids

The objective of this study was to screen wild and domesticated tomatoes for resistance to Tomato yellow leaf curl virus, Israel (TYLCV-Is) and Tomato leaf curl virus from Bangalore isolate 4, India (ToLCV-[Ban4]) to find sources of resistance to both viruses. A total of 34 tomato genotypes resistant/tolerant to TYLCV-Is were screened for resistance to ToLCV-[Ban4] under glasshouse and field conditions at the University of Agricultural Sciences, Bangalore, India. Resistance was assessed by criteria like disease incidence, symptom severity and squash-blot hybridization. All the tomato genotypes inoculated with ToLCV-[Ban4] by the whitefly vector Bemisia tabaci (Gennadius) produced disease symptoms. In some plants of the lines 902 and 910, however, the virus was not detected by hybridization. The tomato genotypes susceptible to ToLCV-[Ban4] by whitefly-mediated inoculation were also found susceptible to the virus under field conditions. However, there were substantial differences between genotypes in disease incidence, spread, symptom severity and crop yield. Despite early disease incidence, many genotypes produced substantially higher yields than the local hybrid, Avinash-2. Sixteen tomato genotypes from India resistant/tolerant to ToLCV-[Ban4] were also tested for TYLCV-Is resistance at the Hebrew University of Jerusalem, Rehovot, Israel. Accessions of wild species, Lycopersicon hirsutum LA 1777 and PI 390659 were the best sources of resistance to both viruses. Lines 902 and 910, which were, resistant to TYLCV-Is were only tolerant to ToLCV-[Ban4] and accession Lycopersicon peruvianum CMV Sel. INRA, resistant to ToLCV-[Ban4], was only tolerant to TYLCV-Is. Implications of using the resistant lines in breeding programme is discussed.     

果树农艺性状基因的分子标记及其应用

总结了果树农艺性状基因标记策略和已获得的分子标记及其在果树上的应用。集团混合分离分析法(BSA)、平行芽变分析法(PSA)和已知信息捕捉法(KIH)是果树农艺性状基因标记筛选的主要方法。目前已获得了大量果树农艺性状基因的分子标记,这些标记是果树基因作图、图位克隆、分子辅助选择和种质资源鉴定等遗传研究与育种实践的有用工具。     

Expression of DNA repair gene ERCC1 and its relationship with PAH-DNA adducts in lung cancer tissues

AIM: To investigate the expression of nucleotide excision repair gene ERCC1 and its relationship with PAH (polycyclic aromatic hydrocarbons)-DNA adducts in lung cancer tissues. METHODS: ERCC1 mRNA expression and the PAH-induced DNA adducts were detected in 150 lung cancer tissues, 120 adjacent lung tissues without cancer cells, 40 benign lung lesions and 40 normal lung tissues. The effects of some exposure factors on the expression of ERCC1 gene and the connection between ERCC1 and PAH-DNA adduct was analyzed. RESULTS: Reduced expression levels of ERCC1 were observed in 46 of 150 (30.7%) lung cancer specimens and 1 of 40 (2.5%) normal lung tissues. Smoking may suppress the expression of ERCC1 gene. The level of PAH-DNA adduct was negatively correlated with the expression of ERCC1 gene, the Spearman coefficient was -0.648, P<0.01. CONCLUSION: ERCC1 is an important nucleotide excision repair gene and may participate in the repair of DNA damage, such as PAH-DNA adduct. Low expression of ERCC1 may play an important role in the development of human lung cancer.