Expression of the mouse metallothionein mutant ββ-cDNA in the lettuces (Lactuca sativa L.)

Mouse metallothionein (MT) domain mutant ββ-cDNA gene has been inserted into the plant expression vector pGPTVd35S which has herbicide (PPT)-resistance gene bar as a selective marker. The chimeric gene was introduced into the lettuce (Lactuca sativa L. cv Salinas 88) by Agrobacterium-mediated transformation. PCR and Southern blot analysis of some putative transformants indicated that the introduced ββ-cDNA was integrated into lettuce genome and inheritted by sexual reproduction. The expression of ββ gene in lettuce plants was demonstrated by Northern and Western blot analysis. Meanwhile, the zinc content of the transformed lettuce plants is up to 400 μg/g dry weight, remarkably higher than the control plants.     

银鲫人工杂交试验中雄核发育现象的RAPD实验证据

以随机扩增多态性ＤＮＡ（ＲＡＰＤ）技术为手段，对生野鲫，雄性异育银鲫及其杂交ＦＩ代（选择性状接近父本的５％Ｆ１代），父本回交Ｆ１你进行了遗传多态分析，从１５０个随机序列引物中筛选得到９种引物，并用它们可得到３６个用作遗传标记的ＲＡＰＤ多态性条带，其中的２２条父本显性条带均遗传子代，而１４条母本性条带则均未被遗传子代，对两亲本和子代中的ＲＡＰＤ标记条带的分布情况及相似系数分析，以及聚类分析结果均支持     

镉对鲫鱼胃肠道嗜银细胞密度的影响

以鲫鱼为研究对象,采用静水染毒法,设对照组和试验组(镉质量浓度0.05 mg/L,0.5 mg/L,5 mg/L)静养10 d,硝酸银染色方法显示鲫鱼胃肠道嗜银细胞,探讨了镉对鲫鱼胃肠道嗜银细胞密度的影响。结果表明,镉对鲫鱼前肠和中肠嗜银细胞数目有明显影响,表现在0.05 mg/L,0.5 mg/L浓度时前肠和中肠嗜银细胞数目明显增加,5 mg/L浓度时则减少;镉对鲫鱼后肠嗜银细胞的影响较小。     

Comparative analysis and evolutionary significance of the HMG1 gene in crucian carp,blunt snout bream,and their polyploid progeny

The full-length mRNA of the high mobility group protein 1 coding gene (HMG1) was obtained by RACE-PCR from red crucian carp (Carassius auratus red var.),blunt snout bream (Megalobrama amblycephala),and their triploid and tetraploid progeny.The sequence contained an open reading frame of 579 nucleotides coding for 193 amino acids.The nucleotide identity of HMG1 was higher between the tetraploid hybrid and the maternal red crucian carp (99%) than between the tetraploid hybrid and the paternal blunt snout bream (97%).The nucleotide identity between the triploid hybrids and each parent (95%) was lower than that between the parents (98%).The protein identity between the tetraploid hybrid and each parent (100%) was higher than that between the triploid hybrid and each parent (97%).Our results suggest that interspecific hybridization generates a shock to the HMG1 gene in triploid hybrids,causing divergence of nucleotides.The HMG1 protein of the tetraploid hybrids was consistent with that of its parents,which reduced the barrier of cross incompatibility between alleles,providing the basis for the bisexual fertile tetraploid hybrids forming a new polyploid species in nature.The secondary and tertiary structures of the HMG1 protein contain eight helices,three switches,two DNA-binding domains in the N-terminus,and a long acidic tail in the C-terminus.Together,these data suggest that the HMG1 protein plays a role of protein-DNA interactions,facilitating various DNA-dependent activities in the nucleus.We also investigated the phylogeny of fish,amphibian,reptilian,bird,and mammalian HMG1 proteins.Our results suggest that HMG1 is an ancestral protein that has been highly conserved.These data provide clues as to how interspecific hybridization may form polyploid hybrids.     

杨树抗杨四瘿螨相关基因的表达分析

采用抑制性消减杂交(suppression subtractive hybridization,SSH)技术,在构建杨四瘿螨诱导差异表达的抑制消减cDNA文库的基础上,通过RTPCR技术对候选基因进行了验证,其中有6条差异表达候选基因在4个时间点表达量有明显的增强。运用RACE(Rapid Amplification of cDNA Ends)技术克隆获得了一段全长2 053 bp的cDNA克隆,命名为PtWRKY,编码588个氨基酸。     

Molecular cloning of growth hormone receptor (GHR) from common carp(Cyprinus carpio L.)and identification of its two forms of mRNA transcripts

Growth hormone receptor (GHR) belongs tothehematopoietic receptor superfamily[1]. The action ofgrowth hormone (GH) in regulating growth[2],re-production[3]and i mmunity[4]has been elucidated.The binding of GHtothe GHRontarget tissues trig-gers a cascade of tyrosine and protein phosphorylationevents, which cul minates in the biological action ofGH[5 ,6]. Up to date GHR cDNAs have been clonedfrom many species[7—9],including various kinds ofmammalian ani mals ; avian of chicken and domest…     

草鱼肌肉生长抑制素cDNA克隆及序列分析

利用RT-PCR技术,从草鱼肌肉组织中克隆出MSTN cDNA序列,长度为1764 bp,编码区为1128 bp,共编码375个氨基酸.前体蛋白包括信号肽序列、N端前肽区、蛋白酶水解位点RIRR及C端活性区(含9个保守的Cys残基).多重序列比较表明,草鱼与斑马鱼、黑头软口鲦之间的MSTN序列同源性在94%以上,与其它鱼类之间的序列同源性也较高(78.1%~82.3%),但与鸟类、哺乳类之间的序列同源性则相对较低.系统进化分析显示,不同物种间的MSTN序列同源性基本反映了它们的亲缘关系.     

鱼类线粒体DNA(mtDNA)及其在分子系统学中的应用

应用线粒体DNA标记,分析种群间的遗传关系、确定种群地理格局、确定种群间的基因流以及确定物种进化显著性保护单元（EsU）,从而为分子系统学研究提供了一个新的具有很强操作性的科学手段。文中论述了鱼类线粒体DNA研究的最新进展,重点介绍了线粒体DNA标记在分子系统学研究中的广泛应用。     

草鱼过氧化物酶体增殖物激活受体(PPAR)基因 cDNA序列的克隆及其组成型表达

过氧化物酶体增殖物激活受体(PPAR)是核激素受体超家族成员之一,分α,β,γ 3个亚型,PPAR具有多种生物学效能,对其基因进行研究将有助于揭示草食性鱼类对营养物利用规律及其机制.通过简并引物克隆草鱼(Ctenopharyngodon idellus)PPAR基因cDNA核心序列,应用3'RACE技术扩增该序列的3'末端序列,并对其进行序列分析,最后用荧光定量PCR方法比较草鱼肝脏、脑、脾脏、肌肉、脂肪和心脏的PPARα、PPARβ和PPARγ基因mRNA相对表达水平.序列分析结果表明,草鱼PPARα、PPARβ和PPARy基因cDNA核心序列片段大小为631 bp,604 bp和651 bp,分别编码210、201和217个氨基酸,PPARy经3'RACE后共获得大小为1 331 bp的片段,编码337个氨基酸.草鱼与鲤鱼(Cyprinus carpio)、斑马鱼(Danio rerio)、鲈鱼(Lateolabrax japonicus)等鱼类PPAR氨基酸序列同源性很高,为71.4%～96.1%;与人(Homo sapiens)、大鼠(Rattus norvegicus)、小鼠(Mus musculus)、牛(Bos taurus)等哺乳动物PPAR氨基酸序列的同源性也较高,为65.0%～77.6%;与非洲爪蟾(Xenopus laevis)等两栖动物PPAR氨基酸序列的同源性约为67.0%.荧光定量PCR结果表明,草鱼PPARα于肝脏,PPARβ于肝脏、肌肉和心脏,PPARγ于肝脏的表达相对较高.相同物种不同PPAR亚型的同源性较低,而不同物种同型PPAR同源性很高,反映了不同亚型在结构上的差异,这与其在功能上的差异是相一致的.     

草鱼脂肪酸去饱和酶和延长酶基因cDNA全序列的克隆与分析

利用RT-PCR和RACE方法克隆得到草鱼(Ctenopharyngodon idellus)肝脏中控制高不饱和脂肪酸(HU-FA)合成的脂肪酸去饱和酶(FAD)和脂肪酸延长酶(ELO)基因cDNA全序列.结果表明,草鱼FAD基因cDNA全长为1 994 bp,开放阅读框(ORF)为1 332 bp,编码444个氨基酸...     

Molecular cloning and polymorphism of major histocompatibility complex class I genes from grass carp (Ctenophayngodon idellus)

In order to clarify the molecular sequences,allelic polymorphism and the tertiary structure of grass carp (Ctenophayngodon idellus) MHC class I,and to further study their relationship with disease resistances,grass carp MHC class I gene (Ctid-MHC I) was cloned from a cDNA library and the allelic polymorphism in the population was investigated.The results showed that most of the variations exist in the peptide-binding domain (PBD) and high polymorphism was identified in the Ctid-MHC I allelic genes from 12 individuals.Based on the genetic distance,Ctid-MHC class I can be classified into 6 types (from Ctid-MHC I-UA to Ctid-MHC I-UF) which were subdivided into 9 lineages (from A to I).Comparison of the Ctid-MHC I among animals and humans showed that the key amino acids of the peptide binding sites are conserved.Analysis of the tertiary structure of the PBD between Grass carp and human crystallographic data of HLA-A2,the variation with insertion or deletion was found in eight regions (A～H).The phylogenetic tree of MHC class I indicates the evolution of MHC class I among grass carp,fish,amphibian,birds,higher vertebrates and humans.     

Effects of 2,3,7,8-TCDD and benzo[a]pyrene on modulating vitellogenin expression in primary culture of crucian carp （Carassius auratus） hepatocytes

Vitellogenin (Vtg) is the precursor of yolk protein. Its expression and secretion are estrogen-regulated and are crucial for oocyte maturation. An in ritro xenoestrogen screening model was established by measuring Vtg induction in cultured primary hepatocytes from crucian carp. Vtg production was detected by biotin-avidin sandwich ELISA method while Vtg and cytochrome P4501A1 (CYP1A1) mRNA induction were measured by semi- quantitative PCR-primer dropping technique. Vtg and Vtg mRNA were dose-dependently induced by diethylstilbestrol (DES, 0.2-200 ng/mL) in hepatocytes of crucian carp. Co-treatment of the DES-induced hepatocytes with either 2,3,7,8-TCDD (TCDD, 0.1-4 pg/mL) or benzo[a]pyrene (B[a]P, 5-1000 ng/mL) resulted in a reduction of Vtg production and an increment of CYP1A1 mRNA expression both in a dose dependent manner, indicating the anti-estrogenic effects of the compounds. However, at lower tested concentrations, TCDD (0.1, 0.2 pg/mL), B[a]P (5 ng/mL)seemed to have a potentiating effect on Vtg expression and secretion, although by their own these compounds had no observable estrogenic effect on Vtg induction. Tamoxifen (a selective estrogen receptor modulators, 1 nmol/L-1μmol/L),and β-naphtho-flavone (β-NF, an aryl hydrocarbon receptor inducing compounds, 2.5-1000 ng/mL) also were employed to study the possible interactions in DES-induced Vtg expression. In co-treatment of the DES-induced hepatocytes with β-NF or tamoxifen, the decrease in Vtg production did parallel induction of CYP1A1 for β-NF, but tamoxifen inhibited Vtg induction did not parallel induced CYP1A1 expression in all test concentrations. On the contrary, it was found that in co-treatment of the TCDD-induced hepatocytes with DES, TCDD induced CYP1A1 mRNA production was inhibited by DES also. These results implicated a possible cross talk between estrogen receptor- and aryl hydrocarbon receptor-mediated pathways in the hepatocytes.     

Effect of enzymatic fish frame protein hydrolysate on freeze-induced denaturation of myofibrillar protein from bighead carp fish （Aristichthys nobilis ） during frozen storage

Three kinds offish frame protein hydrolysates （PPH, APH and FPH） were prepared from fish frame of red drum （ Sciaenops ocellatus ） by papain, alkaline proteinase and flavorzyme treatment. The hydrolysates were mainly composed of peptide （83.5 % -84.6% ） and displayed different molecular weight distribution pattern. The protective effects of hydrolysates on the freeze-induced denaturation of myofibrillar protein （Mf） from bighead carp （Aristichthys nobilis） mince during storage at -20℃ for 12 weeks were investigated. The hydrolysate （5 % dried weight/wet weight） reduced the freeze-induced denaturation of Mf as evidenced by the lowered decrease in Ca-ATPase activity and reactive sulfllydryl contents as well as the impeded increase in surface hydrophobicity. Microscopic photographs indicated that the hydrolysates inhibited the growth of ice crystal in fish mince, and then prevented the aggregation of Mf during frozen storage. The protective effects of hydrolysates on freeze-induced denaturation of Mf were influenced by the molecular weight distribution. PPH had strongest cryoprotective ability among three hydrolysates.     

黑水虻油替代豆油对淇河鲫生长性能、血清生化指标和肠道消化酶活力的影响

为研究黑水虻油替代豆油在淇河鲫生产实践中的效果,以黑水虻油替代淇河鲫基础饲料中不同质量分数的豆油,配制成5种等氮等脂饲料,饲喂初始体质量为(18.40±2.31)g的淇河鲫.56 d后检测其对淇河鲫生长性能、常规组分、血清生化指标、抗氧化能力和肠道消化酶活力的变化.结果显示,各组之间淇河鲫末质量(FBW)、增重率(WG...     

Identification of antiviral- relevant genes in the cultured fish cells induced by UV-inactivated virus

~~Identification of antiviral- relevant genes in the cultured fish cells induced by UV-inactivated virus~~     

Identification of differentially expressed genes in photo-period sensitive genic male sterile rice by randomly amplified cDNAs using RAPD primers

The fertile and sterile young panicle representational populations were constructed by bulked sampling method using young panicles in the photo-period sensitive stage of fertility transformation. Two populations were analyzed using cDNA-RAPD. The results showed that: (i) bulked sampling method can be employed to analyze differentially expressed genes using cDNA-RAPD, taking an advantage in avoiding false positive caused by conventional sampling method. (II) Among 150 random primers used, 83 primers amplified the same banding patterns, and 34 primers amplified the same banding pattern but different staining of intensity on gel. (iii) 33 primers amplified differential cDNA bands between fertile and sterile cDNA populations, and the ratio of polymorphism was 22%. It is concluded that there may exist a lot of genes relating to sterility, which makes the differentially expressed cDNA fragments complicated.     

Identification of differentially expressed genes in photo-period sensitive genie male sterile rice by randomly amplified cDNAs using RAPD primers

The fertile and sterile young panicle representational populations were constructed by bulked sampling method using young panicles in the photo-period sensitive stage of fertility transformation. Two populations were analyzed using cDNA-RAPD. The results showed that: ( i ) bulked sampling method can be employed to analyze differentially expressed genes using cDNA-RAPD, taking an advantage in avoiding false positive caused by conventional sampling method. ( ii ) Among 150 random primers used, 83 primers amplified the same banding patterns, and 34 primers amplified the same banding pattern but different staining of intensity on gel. ( iii ) 33 primers amplified differential cDNA bands between fertile and sterile cDNA populations, and the ratio of polymorphism was 22%. It is concluded that there may exist a lot of genes relating to sterility, which makes the differentially expressed cDNA fragments complicated.