香猪SNP位点rs80894395的多态性及其与体尺指标之间的关联研究

为了探究影响香猪生长发育的功能基因,本试验利用Illumina Porcine SNP60K芯片筛选出N-乙酰氨基半乳糖转移酶2(polypeptide N-acetylgalactosaminyltransferase 2,GALNT2)基因内含子10中一个与腹围呈弱相关的单核苷酸多态性(single nucleotide polymorphism,SNP)位点rs80894395。在此基础上应用等位基因特异性PCR(allele-specific PCR,AS-PCR)技术检测520头雌性香猪群体中该位点的基因型,并与43头大白猪、36头柯乐猪相比较。结果表明,从3个猪群体中检测到rs80894395位点的CC、CT和TT3种基因型;其中香猪群体以CT为优势基因型,且基因型与香猪的体重、体长和腹围呈弱的正相关;大白猪群体以CC基因型为主,T等位基因频率较香猪极显著减少(P<0.01);柯乐猪群体中CC与CT基因型频率相等,与香猪和大白猪的基因频率间的差异不显著(P>0.05)。生物信息学分析表明,rs80894395位点可能影响GALNT2基因的剪接过程。rs80894395与香猪的生长发育可能有一定的联系。     

一个水稻早衰突变体基因的精细定位

【目的】对水稻叶片早衰突变体W330进行遗传分析及精细定位,获得控制突变表型的基因。【方法】用60Co-γ辐射诱变籼型水稻恢复系中恢8015,从突变体库获得一份叶片早衰突变体W330。对该突变体进行表型观察和主要农艺性状调查。利用多代自交稳定的突变体W330与粳稻品种02428杂交,观察F1和 F2的表型,并统计F2群体中早衰突变表型与正常野生型的分离情况,分析该突变表型的遗传行为。利用构建的F2群体进行精细定位和候选基因分析,然后对候选基因进行DNA测序、酶切分析、表达分析、酶活测定及进化分析。【结果】突变体W330从三叶期开始出现叶片衰老,直至抽穗期及黄熟期。与野生型相比,突变体W330株高变矮、分蘖减少、叶片变窄、抽穗期不变、每株有效穗数、每穗着粒数和结实率亦显著降低。W330与02418杂交的F1表现正常,F2群体中正常植株与早衰突变植株的分离比符合3﹕1,表明突变体W330的突变性状受1对隐性核基因控制。利用F2定位群体及SSR、Indel标记,最终将目标基因定位在第3染色体短臂上2个分子标记CD-5与CD-7之间,物理距离约为21.5 kb。基因预测表明该区域共有4个完整的ORFs。其中,LOC_Os03g0131200编码一个过氧化氢酶OsCATC,基因组序列分析表明,W330突变体中的该基因从ATG开始第109位,在第一个内含子的末位发生了一个C到G的颠换,造成第一个内含子没有剪切,最终导致翻译提前终止,酶切试验验证了这一突变位点。与野生型亲本中恢8015相比,W330突变体在三叶期叶片中的过氧化氢酶的活力下降了47.8%,而过氧化氢含量上升了2.7倍。由此,推定W330与OsCATC等位。系统进化分析发现,OsCATC与水稻中同源的过氧化氢酶不在同一进化分支上。实时荧光定量PCR发现,与其野生型相比,突变体W330叶片中的OsCATA和OsCATB的表达量显著上升,而OsCATC的表达量没有明显的变化。推测,这3个高度同源的基因在水稻体内可能存在互补机制。【结论】W330突变体基因是已报道的过氧化氢酶基因OsCATC的等位基因。W330突变体在第一个内含子上发生的一个点突变造成可变剪切的发生,使得水稻一个过氧化氢酶失活,导致突变体W330突变表型的出现。     

K-Domain Splicing Factor OsMADS1 Regulates Open Hull Male Sterility in Rice

We identified the rice floral organ development mutant, termed as open hull and male sterile 1(ohms1), from the progeny of the indica restorer line Zhonghui 8015 treated with 60 Co γ-ray irradiation. The ohms1 mutant exhibited an open hull and lemma- and palea-like structure conversion between the anthers and stigma, which resulted in the ohms1 mutant spikelet showing ‘tridentate lemma'. The ohms1 mutant was entirely sterile but had 60%–70% fertile pollen. Genetic analysis and gene mapping showed that ohms1 was controlled by a single recessive gene, and the mutant gene was fine-mapped to a 42-kb interval on the short arm of chromosome 3 between markers KY2 and KY29. Sequence analysis of the four open reading frames in this region revealed that the mutant carried a single nucleotide transformation(A to G) at the last base of the fifth intron, which was likely corresponded to ohms1 phynotype, in an MIKC type MADS-box gene OsMADS1(LOC_Os03g11614). Enzyme digestion and c DNA sequencing further indicated that the variable splicing was responsible for the deletion of the sixth exon in ohms1, but no structural changes in the MADS domain or amino acid frame shifts appeared. Additionally, real-time fluorescent quantitative PCR analysis showed that the OsMADS1 expression level decreased significantly in the ohms1 mutant. The expression levels of rice flowering factors and floral glume development-related genes also changed significantly. These results demonstrate that OsMADS1 may play an important role in rice floral organ development, particularly in floral glume development and floret primordium differentiation.     

松杉木材无规聚烯烃热熔胶的快速拼接工艺

采用松木和杉木为试验材料,无规聚烯烃(APAO)热熔胶为胶黏剂,以热压温度、压力、进料速度、涂胶量为影响因素,研究工艺参数对木材热熔胶粘接的胶合强度影响。结果表明:热压温度对松木单板间热熔胶胶合性能影响为极显著,热压压力和进料速度对松木单板间胶合的影响为显著;热压温度、热压压力和涂胶量对杉木单板间热熔胶胶合强度的影响均为显著。松木单板间热熔胶胶合的最优工艺为辊压压力0.8 MPa、热熔胶温度150℃、进料速度6 m·min^-1,涂胶量140 g·m^-2,其胶合强度可达1.43 MPa,杉木单板间热熔胶胶合最优工艺为辊压压力0.8 MPa,热熔胶温度160℃、进料速度9 m·min^-1,涂胶量160 g·m^-2,其胶合强度可达1.74 MPa。该木材热熔胶胶拼工艺关键参数的确定,可为木门窗用异型集成材制造及木制品封边自动化应用等提供重要依据。     

肌肉生长发育可变剪接研究进展

为了解动物肌肉生长发育可变剪接调控过程最新研究,本文对近年来人、小鼠和其他多种动物肌肉生长发育的可变剪接调控的相关研究进行了整理与分析,总结了肌肉生长发育可变剪接的发生、调控蛋白多样性的分子机制、人和动物的肌肉发育过程中可变剪接的调控研究进展及可变剪接的数据量化方法.结果 表明:肌肉是表现出最高水平的组织特异性和保守可...     

Biochemical differences in lateral muscle of wild and farmed gilthead sea bream (series Sparus aurata L.)

Red and white muscle from specimens of wild and farmed gilthead sea bream (Sparus aurata) were analyzed for histochemical ATPase activity, total protein content, fatty acids, trace element concentrations and myosin isoforms. The fibre type composition of muscle samples was confirmed histochemically by the ATPase reaction, which did not show any differences between the two groups of animals. Myosin ATPase activities, myosin and protein yields were significantly higher in white muscle than in the red muscle and for the red muscle the latter two parameters were higher in wild fish. Fatty acid profiles revealed differences between the two groups of animals, probably because of the fatty acid composition of the diets. Zinc, copper and iron concentrations were higher in red muscle than in white muscle; muscles from wild fish were significantly richer in trace elements. No separation of fast and slow heavy chains of myosin could be obtained on SDS-gel electrophoresis, but two dimensional electrophoresis revealed the presence of three light chains in white muscle (LC1F, LC2F, LC3F), and two main types in red muscle (LC1S, LC2S). Small, variable percentages of LC3F were found in the red muscle samples, especially in the wild fish. It is concluded that the different environmental conditions, experienced by wild and farmed fish, have significantly influenced the biochemical composition of their lateral muscle.     

植物可变剪接研究进展

选择性剪接广泛存在于植物中,是生物体转录组和蛋白质组多样性的主要来源。随着科技的发展,可变剪接的研究方法逐渐变得简单方便高效,越来越多的可变剪接事件在植物中被发现。本研究对植物可变剪接的机制、研究方法以及几种植物的最新可变剪接研究进展进行分析,并且对未来应深入研究的方向给出了建议。     

高羊茅叶绿体表达载体构建及绿色荧光蛋白基因瞬时表达检测

 【目的】验证高羊茅叶绿体表达载体在高羊茅叶绿体中的瞬时表达情况,为今后在高羊茅叶绿体中稳定表达该载体,获得耐旱高羊茅的叶绿体转基因株系奠定基础。【方法】首先从高羊茅草叶绿体基因组中克隆16S/trnI-trnA/23S片段,作为定点整合同源片段。然后将耐旱相关基因酵母海藻糖合酶基因tps1与水稻叶绿体16S rRNA基因的强启动子Prrn、烟草叶绿体基因psbA的终止子构建表达盒（Prrn-tps1-TpsbA-ter）,连同草丁膦抗性基因bar表达盒、卡那霉素抗性基因nptII和绿色荧光蛋白基因gfp构建的融和基因表达盒一起克隆到定点整合同源片段之间,构建高羊茅叶绿体的稳定表达载体gTKGB。通过基因枪轰击法将gTKGB转化到高羊茅幼嫩叶片中,用激光共聚焦扫描显微镜对GFP的表达情况进行检测分析。【结果】克隆的高羊茅叶绿体基因16S-trnI-trnA-23S在NCBI登录号为：DQ490947－DQ490950。构建的叶绿体表达载体gTKGB转化高羊茅幼嫩叶片,在叶绿体中有很好的GFP表达。【结论】高羊茅叶绿体表达载体gTKGB可用于高羊茅叶绿体转化。