β-甘露聚糖酶的研究现状

综述了β-甘露聚糖酶的作用方式、生物来源、性质、遗传基础及应用。     

植物乙酰辅酶A羧化酶β亚基acc D在因研究进展

综述了乙酰辅酶A羧化酶及其催化域羧基转移酶(Carboxyltransferase,CT)的β亚基(β-CT)的分布和生理功能、β-CT编码基因accD的分子生物学特征及乙酰辅酶A羧化酶在脂肪酸代谢工程中的应用研究进展。     

Identification of Co-dominant SSR Markers Associated with Genes Controlling α′-and α-subunit-null β-conglycinin Phenotypes in Soybean (Glycine max (L.) Merr.)

Studies have shown that the three subunits of β-conglycinin are the main potential allergens of soybean sensitive patients.And β-conglycinin has adverse effects on nutrition and food processing.So solation and production of lines with lower β-conglycinin content has been the focus of recent soybean breeding projects.Soybean lines with deficiency in one or all subunits of β-conglycinin have been obtained.An effective and rapid system to identify such mutations will facilitate genetic manipulation of the β-conglycinin subunit composition.Here,two segregating F_2 populations were developed from crosses between Cgy-1/cgy-1 (CC),an α′-lacking line (Δα′),and DongNong 47 (DN47),a wild-type (Wt) Chinese soybean cultivar with normal globulin components,and Cgy-2/cgy-2 (CB),an α-lacking line (Δα),and DN47.These populations were used to estimate linkage among the cgy-1 (conferring α′-null) and cgy-2 (α-null) loci and simple sequence repeat (SSR) markers.Seven SSR markers (Sat_038,Satt243,Sat_307,Sat_109,Sat_231,Sat_108 and Sat_190) were determined to co-segregate with cgy-1,and six SSR markers (Satt650,Satt671,Sat_418,Sat_170,Satt292 and Sat_324) co-segregated with cgy-2.Linkage maps being composed of seven SSR markers and cgy-1 locus,and six SSR markers and the cgy-2 locus were then constructed.It assigned that the cgy-1 gene to chromosome 10 at a position between Sat_307 and Sat_231,and the cgy-2 gene to chromosome 20 at a position between Satt650 and Satt671.These markers should enable map-based cloning of the cgy-1 and cgy-2 genes.For different subunit-deficiency types[α′-null,α-null and (α′+α)-null types],the two sets of SSR markers could also detect of polymorphism between three normal cultivars and seven related mutant lines.The identification of these markers is great significance to the molecular marker-assisted breeding of soybean β-conglycinin subunits.     

Uterine Expression of WNT7A and β-catenin After Induction of Oestrus in Sheep

WNT7A and β-catenin localisations and roles in regulating periimplantation ovine conceptus development under natural estrous conditions have been elaborated.However,their locations and expression patterns have not been reported under induction of oestrus.The localisation,expression and function of WNT7A and β-catenin in the uterine tissues of the early pregnant and non-pregnant sheep on days 10,12,14,16 and 18 following artificial induction of oestrus were investigated by means of in situ hybridisation,real-time RT-PCR,immuno-histochemistry and western blotting methods.WNT7A and β-catenin mRNA and protein were both restricted to the apical surfaces of the uterine luminal epithelium (LE) and glandular epithelium (GE).In pregnant sheep,protein localisation of WNT7A and β-catenin was observed both in the endometrial LE and GE.Their staining presented on day 10,increased between day 12 and day 16,and decreased on day 18.WNT7A and β-catenin mRNA and protein expression increased initially and then decreased from day 10 to day 18,peaking on day 16,and β-catenin reaching a peak on day 18 in the uterine tissues of pregnant sheep (p0.05).By contrast,no significant changes in WNT7A and β-catenin mRNA and protein expression levels were observed from day 10 to day 18 of the oestrus cycle in the uterine tissues of non-pregnant sheep (p0.05).Additionally,WNT7A and β-catenin mRNA and protein expression levels in the uterine tissues of the early pregnant sheep were significantly higher than those of non-pregnant sheep (p0.05).Treatment of endometrial epithelial cells with WNT7A increased the mRNA expressions of β-catenin,c-myc and Cyclin D1.These results provided an underlying mechanism of periimplantation ovine conceptus development under induction of oestrus.     

富含游离态异黄酮豆豉的制曲工艺优化

为了制备富含游离态异黄酮的豆豉,以前期筛选出的一株高产β-葡萄糖苷酶的菌株为菌种进行制曲。在研究接种量、发酵温度、发酵时间三种单因素对豆曲中蛋白酶和β-葡萄糖苷酶的活力影响基础上,采用三元二次正交旋转组合实验设计对制曲工艺进行优化,确定了最佳的制曲工艺条件为接种量(105.4个孢子·g-1大豆)、制曲温度26.5℃、制曲时间4 d,在此条件下,β-葡萄糖苷酶酶活力为245.24 U·g-1,蛋白酶酶活力为623.17 U·g-1。     

小鼠RPL9基因克隆及组织表达差异性研究

根据已报道的小鼠核糖体蛋白L9基因( RPL9)序列设计PCR引物,克隆RPL9基因,并以β-actin为内参基因,采用相对荧光定量RT-PCR方法,检测、分析RPL9基因mRNA在小鼠心、肝、脾、肺、肾、脑及脂肪7个组织中表达量的差异。结果表明,成功克隆出RPL9基因序列全长,该基因在小鼠的7个组织中均有不同程度的转录表达,其中,在脂肪中的表达量最高,在肾、脾和脑中的表达量较高,在心、肝中的表达量较低,在肺中表达量最低。     

油橄榄叶提取物对海洛因依赖小鼠肺组织结构及IL-1β和TNF-α表达的影响

将100只健康小鼠皮下注射海洛因(10mg/kg)40d,然后分别用不同剂量的(50、100、200mg/kg)油橄榄叶提取物(OLE)进行皮下注射治疗,观察肺脏组织结构的变化,检测肺脏中白介素-1β(interleukin-β,IL-β)和肿瘤坏死因子-α(tumor necrosis factor-A,TNF-α)的表达情况.结果表明:与模型对照组相比,OLE使海洛因依赖小鼠肺泡直径、肺泡隔厚度减小,肺脏中IL-1β和TNF-α的表达明显减弱,且呈较好的量效关系,表明OLE能够对海洛因造成的肺损害起到保护作用,其作用机制可能是OLE抑制了IL-1β和TNF-α的在肺中的表达.     

Identification and quantification of benzimidazole resistance polymorphisms in Haemonchus contortus isolated in Northeastern Brazil

Haemonchus contortus is the most prevalent nematode in Brazil. The objective of this study was to select 6 populations of H. contortus of known or suspected benzimidazole resistance status and characterize these using quantitative real-time polymerase chain reaction (qPCR) for single nucleotide polymorphisms (SNPs) F200Y, F167Y and E198A in the β-tubulin isotype 1 gene. qPCR was performed using DNA from a pool of 10 adult male H. contortus from a single animal per farm. Faecal egg count reduction test (FECRT) and egg hatch test (EHT) were used to determine the resistance status. Samples were obtained from 6 farms located in 5 counties in the Ceará State: Tauá, Boa Viagem, Quixadá, Santa Quitéria and Solonópole. The inbred-susceptible-Edinburgh (ISE) isolate was used as reference for comparative purposes in the qPCR. Benzimidazole resistance was detected by FECRT on all farms with efficacy values ranging from 0 to 51%. EC50 values as determined by EHT were all above 1.49 μg/ml. High frequencies of the resistant SNPs F200Y and F167Y alleles were detected but no resistance was detected at SNP E198A. Our results suggest that the SNPs F167Y and F200Y are both important for benzimidazole resistance in the studied populations.     

糖对长白猪精液NAGaseⅡ的抑制机理

以长白猪精液为材料,采用经典的纯化方法获得N-乙酰-β-D-氨基葡萄糖苷酶（EC 3.3.1.52,NAGase）的2种同工酶,命名为：NAGaseⅠ和NAGaseⅡ。其中NAGaseⅡ的比活力为358.21U/mg,纯化倍数为6.37倍。以海藻糖、D-甘露糖、蔗糖、葡萄糖为效应物,研究其对该酶活性的影响。结果表明：在特定浓度范围内,海藻糖对酶活力基本没有影响;D-甘露糖、蔗糖、葡萄糖对该酶均有一定的抑制作用;D-甘露糖对酶的抑制作用表现为非竞争型,抑制常数KI为11.94mmo/L;蔗糖对酶的抑制作用表现为竞争型,抑制常数KI为0.56mmol/L;葡萄糖对酶的抑制作用表现为混合型,抑制常数KI和KIS分别为0.35mol/L和64.29mmol/L。     

Evaluation of beta-mannanase and nonstarch polysaccharide-degrading enzyme inclusion separately or intermittently in reduced energy diets fed to male broilers on performance parameters and carcass yield

The experimental design consisted of 5 dietary treatments including a positive control (PC), a negative control (NC; with a reduction of 88 kcal/kg of AME through the starter and grower 1 phase and a reduction of 132 kcal/kg of AME in the grower 2, finisher, and withdrawal phases compared with the PC), and the NC supplemented with either β-mannanase, nonstarch polysaccharide-degrading enzymes (NSPase; cocktail carbohydrase), or β-mannanase and NSPase intermittently fed. The intermittent treatment included β-mannanase from d 1 to 21 (starter and grower 1 phases) and NSPase from d 22 to 47. Each treatment included 9 replicate pens with 35 male broilers placed per replicate (1,575 total chicks placed). The dietary program consisted of 5 dietary phases, including the starter through d 10, grower 1 through d 21, grower 2 through d 32, finisher through d 40, and withdrawal through d 47. Broilers were weighed and feed consumption determined on days of dietary changes. On d 48, following an 8-h feed withdrawal, 6 broilers from each replicate pen were removed and processed for whole bird and fat pad measurements. The reduction in energy in the NC diet reduced BW and increased mortality rate, and the inclusion of β-mannanase and NSPase separately and intermittently in the NC diet improved growth performance and reduced mortality to levels that were comparable to the PC. The NC yielded the highest cumulative mortality-corrected FCR and all enzyme inclusion treatments reduced FCR to levels comparable to the PC for the duration of the trial. The NC diet yielded the lowest processing yields and the inclusion of β-mannanase and NSPase separately and intermittently increased multiple processing parameters to a level similar to the PC. These data confirm the ability of β-mannanase and NSPase inclusion separately and intermittently to improve performance parameters in reduced-energy broiler diets.