我国部分地区蛋鸡ALV-J分离株gp85基因遗传进化分析

为了解我国蛋鸡J亚型白血病病毒(ALV-J)株来源及其遗传进化关系,本研究对2009年从我国6个省区蛋鸡场分离到的19株ALV-J的gp85基因进行克隆和测序,并与11个ALV-J参考株gp85基因作了比较分析.结果表明:19个ALV-J分离株的gp85基因长度为894bp～924 bp不等,分别编码298～308个氨基酸;各病毒株间gp85推导氨基酸的同源性为71.3%～100%.遗传进化分析表明,目前我国蛋鸡ALV-J分离株来源复杂,其中13个分离株与英国原型株HPRS-103、国内麻黄肉鸡株SCAU-0901亲缘关系较近;3个分离株与美国株ADOL-7501及国内白羽肉鸡株HN0001处在同一大的分支;而另外3个分离株则各自形成独立的分支,表明其gp85基因发生了较大变异.本研究表明,19个分离株与国内早期肉鸡分离株亲缘关系较远,提示我国当前蛋鸡ALV-J株可能并非源自国内早期肉鸡ALV-J株,其来源有待进一步研究.     

玉米品种‘新科19’花期调节制种技术的研究

花期相遇良好是确保制种成功的关建,通过进行玉米杂交种‘新科19’双亲的播期试验及雌、雄穗分化观察,掌握了2个自交系的穗分化及吐丝、散粉规律,明确了‘新科19’正交制种播期调节内在原因及花期预测的叶龄指标。     

油用向日葵新葵19号恢复系654R的选育及利用

[目的]研究油用向日葵恢复系的选育方法。[方法]在现有技术中向日葵恢复系的选择过程比较漫长且结果不明确的情况下,为了改变从杂种一代F1以后连续自交的作法,在早代就开始进行测定其配合力,有目的的选择,方向比较明确,提高选择效率,创造新的优异资源。[结果]选育出油用向日葵新葵19号恢复系654R,该恢复系植株性状好、抗病性好、抗逆性强、配合力高、杂种优势强。[结论]该研究提供了一种油用向日葵恢复系新的选育方法,并对654R进行利用。     

25％氰烯菌酯悬浮剂防治水稻恶苗病的田间药效

[目的]明确新型杀菌剂25%氰烯菌酯悬浮剂对水稻恶苗病的防治效果,为黑龙江省水稻生产中有效防治恶苗病提供参考。[方法]采用田间药效试验方法,评价25%氰烯菌酯悬浮剂对水稻的安全性及对恶苗病的防治效果。[结果]25%氰烯菌酯悬浮剂0.25ml/kg种子或25%氰烯菌酯悬浮剂0.15 ml/kg种子与种衣剂混用对水稻安全且对水稻恶苗病防治效果较好,明显好于对照药剂咪鲜胺和单独使用种衣剂。[结论]建议在黑龙江省水稻生产中推广应用25%氰烯菌酯悬浮剂。     

伪狂犬病病毒Fa株gp63（gI） 基因核苷酸序列测定及分析

对我国最早分离的伪犬病病毒（pseudorabies virus,PRV)Fa株糖蛋白gp63(gI)基因核苷酸序列及的氨基酸序列作了测定与分析，完整的gp63(gI)结构基因从起始密码子ATG到终止密码子TGA共有1050个核苷酸残基，编码由439个氨基酸残基构成的多肽链，4种核苷酸残基的构成比分别为：G35．9％，11.33%,T11.81%,C40.95%,G C含量达76．85％，PRV Fa株gp63基因核苷酸序列与Nice株的相比，两者同源性达98．13％，仅存在3个核苷酸残基的缺失变异，氨基酸推导序列分析表明，糖蛋白gp63具有典型膜蛋白特征，与Nice株相比，同源性为97．42％，gp63基因起始密码子由3个连续的ATG组成，ATG上游区域内无启动子调控区序列。     

Construction of siRNA-survivin and GRIM-19 coexpression plasmid and its growth inhibitory effect on prostatic cancer DU145 cells

AIM: To construct the recombinant plasmid that expresses siRNA-survivin and GRIM-19 simultaneously, and to identify the validity of the recombinant plasmid and observe its effect on expression of survivin and GRIM-19 and proliferation ability of prostate cancer DU145 cells. METHODS: The recombinant plasmid coexpressing siRNA-survivin and GRIM-19 was constructed using gene cloning technique. The prostatic cancer DU145 cells were transfected with the coexpression plasmid and control plasmids. Survivin and GRIM-19 mRNA expression was detected by semi-quantitative RT-PCR. The proliferation ability affected by coexpression plasmid was measured by MTT assay. RESULTS: The coexpression plasmid pGRIM-19-si-survivin was successfully constructed according to DNA recombinant technique and identified through restriction enzyme digestion and plasmid sequencing. Compared with the mock, survivin mRNA expression levels were 0.55?0.05,0.62?0.08 and 0.35?0.05 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively. Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin inhibited survivin mRNA expression markedly (P<0.05), while the expression levels of GRIM-19 mRNA were 1.93?0.14, 2.57?0.20 and 4.12?0.21 in psi-survivin, pGRIM-19 and pGRIM-19-si-survivin groups, respectively (P<0.01). Compared with pGRIM-19 group, pGRIM-19-si-survivin enhanced GRIM-19 mRNA expression more obviously (P<0.05). After transfection for 48 h, the proliferation rates were 58.0%?7.2%, 62.1%?6.1% and 50.2%?4.8% in the 3 experiment groups compared with the mock (P<0.05). After transfection for 72 h, the proliferation rate were 43.4%?4.3%, 51.3%?6.7% and 26.8%?7.1% in experiment groups compared with the mock (P<0.05). Compared with psi-survivin and pGRIM-19, pGRIM-19-si-survivin significantly inhibited the cell growth (P<0.05). CONCLUSION: Transfection of coexpression plasmid pGRIM-19-si-survivin dramatically changes the mRNA expression of survivin and GRIM-19 and inhibits the cell growth.     

Morphological study of macrophages infected with constructed recombinant adenovirus carrying gp120 gene of Chinese HIV-1 strain

AIM: To construct a recombinant adenovirus carrying gp120 gene of Chinese HIV-1 strain,which can infect mouse bone marrow-derived macrophages (BMM). METHODS: Co-transfection of shuttle and backbone plasmids of AdMax system into 293Ad5⁺ cells was performed, followed by viral packaging, propagation and purification. These viruses were subject to Karber TCID₅₀ titration. The expression of gp120 protein in 293Ad5⁺ cells was determined by ELISA. The viral titration was validated by a multiplicity of infection (MOI) test with BMM. RESULTS: The titers of the outcome viruses, including AdMax-HIV-1 gp120 (Ad-gp120) and its vector control Ad-GFP, were 10^8.3 and 10^8.1 TCID₅₀/mL, respectively. Both recombinant adenoviruses infected BMM with similar capacity of 293Ad5⁺ cell infection, which validated the TCID₅₀ titration.The gp120 protein was positive in 293Ad5⁺ cell lysates. BMM activation was observed morphologically after Ad-gp120 infection as compared with Ad-GFP-infected cells. CONCLUSION: Functional adenovirus containing HIV-1 gp120 of prevalent strains in China was successfully constructed. Infection of Ad-gp120 causes BMM activation.     

Antinociceptive and antiplasmodial activities of cassane furanoditerpenes from Caesalpinia volkensii H. root bark

The chloroform and ethyl acetate extract (100 mg/kg) of Caesalpinia volkensii H. exhibited significant (P ≤ 0.05) antinociceptive activities using hot plate and writhing tests in mice while the later showed antiplasmodial activity (IC₅₀ 0.23 ± 0.07 and 4.39 ± 2.49 μg/ml) against chloroquine sensitive (D6) and chloroquine-resistant (W2), respectively. Two new furanoditerpenes [rel. 1β,5α-dihydroxyvoucapane (1) and rel. 1β,6β-dihydroxyvoucapane; 19β-methyl ester (2)] together with seven known compounds [voucapane (3), voucapan-5-ol (4), deoxycaesaldekarin C (5), caesaldekarin C (6), 5-hydroxyvinhaticoic acid (7), triacontanyl-(E)-ferulate (8), triacontanyl-(E)-caffaete (9) and 30′-hydroxytriacontanyl-(E)-ferulate (10)] were isolated from the two extracts. The administration of 3, 4, 5 and 6 (100 mg/kg i.p) caused a significant (P ≤ 0.05) reduction in the number of writhing episodes induced by acetic acid and (P ≤ 0.01) increased pain latency threshold in hot-plate test compared to control. However, the pure compounds indicated relatively (P ≤ 0.05) low antiplasmodial activity. The phytochemical constituents from the root bark of C. volkensii had better analgesic properties than antimalarial properties, justifying the use of the plant root bark as a remedy for pain.     

华南型黄瓜新品种吉杂19的选育

吉杂19是以自交系04196为母本,以自交系Ⅱ-0407-1-2-1-1为父本配制而成的保护地华南型黄瓜一代杂种.早熟,第1雌花节位为第4节左右,播种至始收60 d(天)左右;植株生长势较强,主侧蔓均可结瓜,结成性好;瓜条棒状、顺直,刺瘤稀少,瓜皮嫩绿色,瓜长22 cm左右,横径4 cm左右,平均单瓜质量150 g;田...     

一株J亚型禽白血病病毒的分离鉴定及其gp85基因的序列分析

为确定安徽巢湖某地方品种养鸡场的疑似禽白血病(avian leukosis,AL)的病原,以DF-1细胞从肿瘤组织中分离病毒,采用RT-PCR方法进行鉴定,并对其感染细胞上清进行透射电镜观察。同时对送检病鸡的心、肝、脾及腺胃等进行组织病理学观察,最后将扩增出的gp85基因与GenBank收录的其他ALV序列进行比对分析。结果表明:PCR扩增产物大小约为928 bp,经测序证实为ALV gp85基因,将该分离株命名为AHCH-2018株。透射电镜观察到具有囊膜的大小约100 nm的病毒粒子。组织病理学结果显示,病鸡的心、肝、脾及腺胃中有大量成灶淋巴细胞增殖。遗传进化分析结果表明,分离株的gp85基因与多株ALV J参考毒株核苷酸序列的同源性为94.1%~99.5%,且与GD1401J株的核苷酸同源性最高,达99.5%,与A、B、C、D、E亚群同源性为46.5%~49.7%。该分离株与J亚群原型毒株处于同一大分支。本研究为了解安徽土鸡种群中禽白血病的分子流行病学特点提供了一定的数据参考。