苜蓿银纹夜蛾核多角体病毒在家蚕蛹体内的复制及重组病毒外源基因的表达

家蚕蛹在复眼着色期,经4℃冷藏24小时后,可被Ac NPV(苜蓿银纹夜蛾核多角体病毒)感染。在蛹体内复制出的多角体大小差异较大,且比在昆虫Sf—21细胞中繁殖的Ac NPV和在蚕蛹体内形成的Bm NPV多角体小。在蛹体内繁殖的Ac NPV的游离病毒可以回返感染Sf—21细胞。由蚕蛹内分离的Ac NPV基因组DNA的限制性内切酶图谱与野生型的Ac NPV相同。以上结果证明Ac NPV可以在蚕蛹体内复制。含HBsAg基因(人乙肝表面抗原)的重组Ac NPV亦可感染家蚕蛹,并能正确表达外源基因。此外,还发现化蛹后第2天的家蚕蛹被注射约5×10~5PFU的Ac NPV,可诱导“人工滞育蛹”现象的发生,在25℃经过30天后蛹仍存活,发育停滞在复眼着色前阶段。     

真空渗入法转pinⅡ基因菜薹外源基因的遗传与表达

 本研究对利用真空渗入法获得的转bar基因及pinⅡ基因菜薹，进行了外源基因的遗传及表达情况分析。结果表明通过该方法获得的转基因植株，外源基因能够稳定遗传。bar基因在 代的分离大部分符合孟德尔遗传规律，但也有不符合的群体；pin II基因在 代中稳定表达，但各个株系之间的表达水平差异较大，因而各个株系的抗虫效果也具有显著差异，即使具有相同外源基因拷贝数的各个株系，其PIN I／蛋白表达量和抗虫效果也具有显著差异。     

关于修订桃种质资源(Prunus persica)描述体系的建议

评价是种质资源工作的重要环节之一,是资源利用与共享的基础。目前,中国桃种质资源的评价描述体系一般是按照中国1990年编制的“桃种质描述符”进行。通过对国际植物遗传资源委员会1980年编制“桃描述系统″和中国“桃种质描述符”在评价的层次、评价内容、评价方法和评价标准的系统比较,以及2个标准在种质资源科研实践使用过程中存在的不足之处,结合中国桃种质资源工作的特点和国外资源的描述方法的新进展,提出中国桃种质资源描述系统的修改建议,为建立中国标准化、数字化的桃种质资源描述系统奠定基础。     

Effects of blocking the expression of PKARⅠβ gene with antisense nucleotide on Shuang long-Jie gu Pill (SLJGP) containing serum osteoblast proliferation

AIM: To further investigate the role of PKARⅠβ in the growth-promoting effects of shuang long Jiegu pill (SLJGP), a Chinese medicine, on cultured osteoblasts. METHODS: pcDNA- antiPKARⅠβ, a recombinant expressing the antisense sequence of PKARⅠβ, was constructed and transformed HFOB1.19 by lipofectin. MTT was undertaken to assess the cell growth with the treatment of high dosage of SLJGP containing serum. RESULTS: Antisense gene blocked the growth-promoting effects of SLJGP containing serum on HFOB1.19. CONCLUSION: The function of SLJGP is closely related to cAMP-dependent protein kinase A.     

猪胸膜肺炎放线杆菌外膜蛋白(OMP)5′末端保守区基因的克隆、表达及其免疫活性研究

以猪胸膜肺炎放线杆菌血清7型25-4株基因组DNA为模板,PCR方法扩增外膜蛋白(OMP)5′末端保守区基因片段(OMPc),酶切及核苷酸序列分析鉴定后,与原核表达载体质粒pGEX-6P-1进行连接,构建成重组表达载体pGEX-OM-Pc,转入大肠杆菌BL21中,以IPTG进行诱导,SDS-PAGE电泳分析发现,转化了重组质粒的菌株所表达的融合蛋白相对分子量为34 kD,与实际预测相符,命名为GST-OMPc.GST亲和层析柱进行纯化,ELISA方法对纯化蛋白进行检测.结果表明:纯化蛋白GST-OMPc能够与兔抗猪胸膜肺炎放线杆菌血清7型的阳性血清反应.OMPc蛋白的成功表达为其功能的研究打下基础.     

透明颤菌血红蛋白基因(vgb)在大肠杆菌中的高效表达

利用PCR技术将透明颠菌血红蛋白基因（vgb）克隆到融合表达载体pET28a，在大肠杆菌BL21（DE3）表达。重组蛋白在30℃诱导获得可溶表达。利用Ni^2＋亲和层析对重组蛋白进行了纯化，得到重组的透明颠菌血红蛋白，蛋白呈红色。实验现象显示重组蛋白与血红索相结合。紫外光区和可见光区波长扫描分析显示：蛋白粗提物和纯化后的血红蛋白在230nm和413nm都有强吸收峰，初步表明，尽管重组蛋白比天然蛋白多36个氨基酸，重组蛋白仍然具有生理活性。诱导后4h和6h的培养液氨基乙酰丙酸含量分别达到17mg／L和21mg／L。说明重组蛋白的表达明显促进了体内heme合成途径。     

凤头白鸭与连城白鸭的群体遗传多样性分析

为系统地评价、保护凤头白鸭资源的遗传多样性,进一步对其开发利用,本研究利用12个微卫星位点(STR分型技术)对凤头白鸭与连城白鸭(各60只)2个群体的遗传多样性参数进行检测和计算。结果表明:除位点AJ272578以外,凤头白鸭群体在APL2、AJ272577、AJ272579、AJ515884、APH01、AY493256、AY4932898、CMO11等8个STR位点上的等位基因多于连城白鸭,在AJ515887、AJ515893、AY493313等3个位点上与连城白鸭相等;凤头白鸭群体的有效等位基因数(Ne)、期望杂合度(He)、观察杂合度(Ho)、多态信息含量(PIC)均大于连城白鸭;2个群体的群体内近交系数(Fis)分别为-0.02、0.09,凤头白鸭群体表现为杂合子过剩。综上,凤头白鸭群体的遗传多样性较连城白鸭更为丰富。     

狼山鸡保种群不同世代MHC BF基因遗传多样性研究

通过Illumina平台Miseq重测序方法分析不同世代(G0、G5、G10、G12、G14、G15、G17)的MHC BF1和BF2基因座位多态性,揭示狼山鸡不同世代间MHC BF基因遗传多样性,为更好地监测保种效果提供科学的决策依据。在狼山鸡7个世代中分别鉴定出MHC BF1、BF2基因59和34个SNPs。狼山鸡MHC BF1基因观察杂合度(Ho)、期望杂合度(He)和多态信息含量(PIC)在G10、G12、G14保持稳定,Ho在不同世代变化情况为G17G15G14、G12、G10G5G0,He和PIC在不同世代变化情况为G17、G15G14、G12、G10G5G0,MHC BF2基因的Ho在G10、G14、G15保持稳定,其他世代变化情况为G17、G12G15、G14、G10G0、G5;He和PIC在G12、G14、G15和G17保持稳定,在其他世代变化情况为G17、G15、G14、G12G10G5、G0。从本研究结果看,狼山鸡不同世代MHC BF1和BF2遗传多样性变化规律不太一致,但总体规律符合波动选择(时空变换的选择)假说,随着时间的推移MHC BF基因遗传多样性变得更为丰富,在某些连续几个世代群体遗传多样性基本保持稳定,这可能与MHC BF基因功能有关。     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.