不同浓度CO2对牛卵母细胞体外成熟和受精的影响

对牛卵巢卵泡内卵母细胞在不同浓度的CO2培养箱内体外成熟、体外受精后的卵裂率及胚胎体外发育进行了研究。结果显示,含5%小牛血清(CS)的TCM-199内的卵母细胞在2%CO2条件下培养20h后,达到第二次减数分裂期(MⅡ期)的卵子数明显高于在5%CO2条件下培养的卵母细胞数(P<0.05)。两种浓度的CO2条件下培养的牛卵母细胞体外受精后的卵裂率无明显差异,2%CO2浓度下培养的卵母细胞的囊胚发育率高于5%CO2浓度下培养的卵母细胞。     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

由Cu(Ⅱ)促进的肌红蛋白选择性水解切割反应

采用聚丙烯酰胺凝胶电泳技术,研究了中性条件下[CuL(Cl)](C lO4)与马心肌红蛋白的选择性水解切割反应,L为2-[二(2-氨乙酸)氨基]乙醇.结果表明,[CuL(Cl)](ClO4)对马心肌红蛋白显示出较高的切割选择性和切割效率,且切割效率与浓度、介质条件、温度、温育时间有一定的相关性.     

Intratree Variability of Cleavage Resistance of Chinese Fir from Plantation

INTRODUCTION Chinese fir (Cunninghamia lanceolata) is a very important timber species in the southern China (Shi Jisen et al. 1987). It is good, fast-growing, and exempts from serious plant diseases. Its plantation is big in terms of size in the South of China and its storage stock is very rich, which is about 20% of the national commodity wood (Zhou Shuangquan et al. 2000). And the lumber becomes mainly commercial construction materials in China because of straight grain, light and so…     

四种泡桐材抗劈力研究

摘要：本文对兰考泡桐、秋叶泡桐、白花泡桐、毛泡桐的径、弦面抗劈力进行了对比研究。结果表明：兰考泡桐、秋叶泡桐、白花泡桐、毛泡桐的径面、弦面抗劈力均值均不相等,其中以白花泡桐值最大,径面抗劈力均值为13．62N／mm,弦面抗劈力均值为13．97N／mm;兰考泡桐最小,径面抗劈力均值为10．81N／mm,弦面抗劈力均值为10．59N／mm。方差分析结果表明：4种泡桐种间,径面抗劈力和弦面抗劈力均有极显著差异;种内,径面抗劈力与弦面抗劈力比较,无显著差异。     

牛不同直径卵泡内卵母细胞体外受精后的卵裂率和染色体异常分析

将不同直径牛卵泡内卵母细胞体外成熟培养22h后，对其体外受精后的卵裂率，胚胎发育速度和胚胎染色体异常发生率进行了研究。结果表明，体外受精后48h，直径3－6mm卵泡内卵母细胞外受精后的卵裂率明显高于直径1－2mm和7－10mm卵泡内卵母细胞体外受精后的卵裂率（P＜0．05）。体外受精后48h，直径3－6mm包泡内卵母细胞体外受精后的胚胎的发育速度明显快于直径1－2mm和7－10mm卵泡组胚胎（P＜0．05）。通过对不同卵裂期胚胎的染色体标本分析表明，直径1－2mm卵泡内卵母细胞体外受精后的胚胎染色体异常发生率明显高于直径3－6mm卵泡组（P＜0．05），但与直径7－10mm卵泡组之间无显著性差异。     

NDV融合蛋白裂解位点基因的重组昆虫杆状病毒表达载体的构建与鉴定

将新城疫病毒（ＮＤＶ）Ｆ４６Ｅ９株和ＬａＳｏｔａＥ４株的融合蛋白裂解位点（Ｆｃ）基因从本实验室构建的重组质粒ｐＵＣＦ４６Ｆｃ和ｐＵＣＬａＦｃ中用ＥｃｏＲＩ和ＳａｌⅠ切出。将这２个毒株的Ｆｃ基因定向克隆入昆虫杆状病毒表达载体ｐＳＸＩＶＶＩ＋Ｘ３的ＥｃｏＲＩ和ＳａｌⅠ位点之间，获得２个毒株的Ｆｃ基因克隆ｐＸ３Ｆ４６Ｆｃ和ｐＸ３ＬａＦｃ。重组质粒用ＥｃｏＲＩ／ＳａｌⅠ、ＰｓｔⅠ酶切法、ＰＣＲ和核酸探针法进行了鉴定，证明插入的基因来自ｐＵＣＦ４６Ｆｃ和ｐＵ－ＣＬａＦｃ，插入的位置和方向都正确。这２个载体的构建为Ｆｃ基因在昆虫细胞系统的表达创造了条件。     

Deduced Amino Acid Sequences Surrounding the Fusion Glycoprotein Cleavage Site and of the Carboxyl-terminus of Haemagglutinin–Neuraminidase Protein of the Avirulent Thermostable Vaccine Strain I-2 of <Emphasis Type="Italic">Newcastle disease virus</Emphasis>

A single-tube RT-PCR technique generated a 387 bp or 300 bp cDNA amplicon covering the F₀ cleavage site or the carboxyl (C)-terminus of the HN gene, respectively, of Newcastle disease virus (NDV) strain I-2. Sequence analysis was used to deduce the amino acid sequences of the cleavage site of F protein and the C-terminus of HN protein, which were then compared with sequences for other NDV strains. The cleavage site of NDV strain I-2 had a sequence motif of ¹¹² RKQGRLIG¹¹⁹, consistent with an avirulent phenotype. Nucleotide sequencing and deduction of amino acids at the C-terminus of HN revealed that strain I-2 had a 7-amino-acid extension (VEILKDGVREARSSR. This differs from the virulent viruses that caused outbreaks of Newcastle disease in Australia in the 1930s and 1990s, which have HN extensions of 0 and 9 amino acids, respectively. Amino acid sequence analyses of the F and HN genes of strain I-2 confirmed its avirulent nature and its Australian origin.