以HSV-1扩增子载体构建的新型重组AAV载体包装系统

将缺失两端ITR的AAV -2基因组克隆于HSV -1扩增子载体获得了质粒pHSV AAV .以HSV- 1tsK株为辅助病毒感染转染了该质粒的Vero细胞 ,可获得一种具有包装重组AAV质粒能力的混合毒种 .连续传代可使其包装能力迅速升高 ,至 5～ 6代达到最高并趋于平稳 .此混合毒种中 ,含有包装了多拷贝AAV -2基因组的复制缺陷性假型HSV- 1病毒 ,它可表达AAV -2编码的复制与包装所需的各种反式蛋白 ;同时也含有HSV -1tsK病毒 ,它提供AAV -2复制与包装所需的全部辅助病毒功能 .实验结果也表明 ,HSV -1tsK病毒的复制为混合毒种辅助重组AAV载体的包装所必需 .这项研究首次报道了一种全新的重组AAV载体包装系统 ,与传统的双质粒共转染、腺病毒辅助包装方案相比 ,本系统简便易行 ,并明显提高了重组AAV的包装效率 .     

Molecular characterization and functional expression of opioid receptor-libe_1 receptor

INTRODUCTIONOpioidactsinthecentralandperipheralnervoussystem(CNSandPNS)to'producenumerouspharmacologicaleffects.Repetitiveorcontinuoususeofopioids,however,causesdrugtoleranceanddependence.Threesubtypesofopioidreceptors,termedp)6,andK,havebeencloned.TheyarecoupledtotheinhibitoryGproteinandnegativelyregulateadenylylcyclasell].RecentlyanewG-proteincoupledreceptortermedopioidreceptor-likereceptor(ORLI)whichbelongstothenewlycharacterizedopioidreceptorsubfamilyhadbeenclonedfromhumanbrainst…     

PERFORMANCE,CARCASS CHARACTERISTICS,MEAT QUALITY AND PLASMA CONSTITUENTS OF MEAT TYPE DRAKES FED DIETS CONTAINING DIFFERENT LEVELS OF LYSINE WITH OR WITHOUT A MICROBIAL PHYTASE

This experiment was conducted to study growth performance, carcass characteristics, meat quality and plasma constituents of Campbell drakes fed diets containing different levels of lysine with or without a microbial phytase. Basal vegetable duck all-mash diets were fed during the growing (1- 35 d of age), and finishing period (36 - 56 d of age) and were formulated to contain 0.90% and 0.73% lysine (negative control), respectively. These diets were supplemented or not with L-lysine HCl, which resulted in a dietary lysine level of 0.90, 0.95, 1.01 and 1.06% and 0.73, 0.80, 0.87 and 0.94%, during the growing and finishing period, respectively. Furthermore, the diets were fed with or without 600 FTU phytase (Natuphos®) except for those containing 1.06 and 0.94% lysine during the growing and finishing period, respectively (positive control). A lysine level of 1.01/0.87% in the growing/finishing diet significantly increased BWG and improved FCR of drakes by 2.1 and 1.8%, respectively. Phytase significantly increased BWG by 2.1% and 3.5% after feeding the basal diet and 1.01/0.87% lysine, respectively. Also, FCR was significantly improved by 2.2 and 1.8% of groups fed 0.95/0.80, and 1.01/0.87% lysine, respectively. Phytase as an independent variable increased BWG by 1.8, and improved FCR by 1.0%. Lysine and/or phytase did not affect carcass yield, and meat quality treats as well as plasma constituents of drakes. However, lysine level at 0.95/0.80% and 1.01/0.87% significantly decreased abdominal fat deposition compared to either the negative or the positive control. In conclusion, a lysine level of 1.01/0.87% in the growing/finishing diets for drakes is adequate. After phytase supplementation of the basal diet the BWG at a lysine level of 0.90/0.73% were similar to the positive control (1.06/0.94% lysine). However, the best FCR was obtained after feeding diets containing 1.01/0.87% lysine supplemented with phytase.     

KILLER WHALE (ORCINUS ORCA) AUDITORY EVOKED POTENTIALS TO RHYTHMIC CLICKS

Temporal auditory mechanisms were measured in killer whales ( Orcinus orca ) by recording auditory evoked potentials (AEPs) to clicks. Clicks were presented at rates from 10/sec to 1,600/sec. At low rates, clicks evoked an AEP similar to the auditory brainstem response (ABR) of other odontocetes; however, peak latencies of the main waves were 3–3.7 msec longer than in bottlenose dolphins. Fourier analysis of the ABR showed a prominent peak at 300–400 Hz and a smaller one at 800–1,200 Hz. High-rate click presentation (more than 100/sec) evoked a rate-following response (RFR). The RFR amplitude depended little on rate up to 400/sec, decreased at higher rates and became undetectable at 1,120/sec. Fourier analysis showed that RFR fundamental amplitude dependence on frequency closely resembled the ABR spectrum. The fundamental could follow clicks to around 1,000/sec, although higher harmonics of lower rates could arise at frequencies as high as 1,200 Hz. Both RFR fundamental phase dependence on frequency and the response lag after a click train indicated an RFR group delay of around 7.5 msec. This corresponds to the latency of ABR waves PIII-NIV, which indicates the RFR originates as a rhythmic, overlapping ABR sequence. The data suggest the killer whale auditory system can follow high click rates, an ability that may have been selected for as a function of high-frequency hearing and the use of rapid clicks in echolocation.     

涠洲岛海域球形棕囊藻藻华过程中营养盐及浮游植物群落组成分析

球形棕囊藻(Phaeocystis globosa)是近年来北部湾常见的有害藻华原因种,涠洲岛海域作为广西沿海藻华的高发区之一,目前仍缺乏关于该海域球形棕囊藻藻华过程的相关研究。为探究涠洲岛海域球形棕囊藻藻华过程中营养盐的状况及藻华的发生对浮游植物群落组成的影响,对2017年3月下旬涠洲岛周边发生的球形棕囊藻藻华区域进行取样调查。结果表明,藻华期间水体中溶解有机磷含量与细胞及囊体密度呈显著正相关。本次藻华主要受水体中溶解态无机氮含量影响,多个调查站点的无机氮/无机磷(DIN/DIP)<10,且硅酸盐/无机氮(SiO₃^2–/DIN)>1,表明这些站点存在氮限制现象,而氮限制可能是本次藻华消亡的主要因素。调查区域共鉴定出浮游植物4门58种,球形棕囊藻细胞密度最高可达1.04×10⁸ cells/L,占浮游植物总细胞密度的98.28%。球形棕囊藻细胞密度与物种多样性指数存在显著的负相关关系。在营养缺乏条件下,有利于球形棕囊藻成囊的链状硅藻种类具有相对较大的细胞密度。     

高危型HPV和L1壳蛋白与宫颈病变关系的研究进展

宫颈病变是女性最常见的疾病之一,是多种因素共同作用的结果。宫颈癌及癌前病变的发生、发展是一个多因素、多步骤的复杂过程,多种基因的改变引发细胞的增殖失控。大量资料表明高危型人乳头瘤病毒(HPV)感染是宫颈病变的主要危险因素,其中HPV16、18型感染占了绝大部分。HPV整合状态与宫颈病变程度密切相关,人乳头瘤病毒(HPV)L1壳蛋白为免疫杀伤HPV病毒的主要靶位,L1蛋白的表达有利于激发人体细胞免疫,清除感染的细胞。HPV感染机体且病毒处于复制阶段时L蛋白才在机体中表达,但是当HPV病毒DNA与宿主细胞基因整合后,L1壳蛋白将不表达,无法形成一系列免疫反应,引发宫颈癌。HPV L1壳蛋白的表达缺失与宫颈病变的进展密切相关。本文对高危型HPV与HPV L1壳蛋白在宫颈病变中的研究进展作一综述。     

低能离子束促进小麦copia-反转录转座子的转录与转座

LTR-反转录转座子是真核基因组重要的组成部分,能通过RNA中间体完成在基因组上的转座。小麦种子受低能氮离子束注入后能促进其反转录转座子的转录,经不同剂量的氮离子束注入的种子发芽24h和48h后,其中的copia-反转录转座子的转录都有不同程度的提高,最高可达到对照的40倍。用基于反转录转座子扩增多态性和反转录转座子微卫星扩增多态性2种分子标记技术扩增经低能氮离子注入后的小麦DNA,指纹图谱显示出了一定的多态性,这说明低能氮离子束激活了小麦中反转录转座子的转座活性。转座子转录活性的增强可调节其邻近基因的表达和mRNA的拼接,反转录转座子插入到基因组上新的位点后,使基因组上的基因发生了重排,这种重排可能表现为植物表型的改变。因此,推测反转录转座子的转录活性提高和转座激活是产生低能离子束注入生物效应的重要原因之一。