杂交

Methods Monoclonal antibody against the glycophorin A was used to identify nucleated erythrocytes in the peripheral blood of pregnant women.Candidates fetal cells from 3 pregnancies at 16 to 24 weeks of gestation were isolated successfully.Sorted cells were also confirmed using Y-special probe (PY3.4) repeated sequences in maternal blood from women carrying male fetuses were detected by polymerase chain reaction.Results The PY3.4 position hybridization rates were 31.5%,0 and 38% respectively in sorted GPA positive cells of 3 samples by FISH,and the results of GPA+ cells of the 3 cases by PCR were positive,negative and positive respectively,which were manifested by the gender after aborticide.

选用红细胞特异单克隆抗体Glycophorin A结合流式细胞术成功地从3例妊娠16～24周的孕妇血中分离出NRBCs，并用Y-染色体特异重复序列DNA探针(PY3.4)荧光原位杂交及聚合酶链反应技术检测Y-染色体特异序列结果 FISH结果，3例分选出的GPA阳性细胞中PY3.4杂交阳性率分别是31.4%、0、38%；其PCR结果，分别为阳性、阴性、阳性，并在引产后得到性别证实。

Methods Monoclonal antibody against the glycophorin A was used to identify nucleated erythrocytes in the peripheral blood of pregnant women.Candidates fetal cells from 3 pregnancies at 16 to 24 weeks of gestation were isolated successfully.Sorted cells were also confirmed using Y-special probe (PY3.4) repeated sequences in maternal blood from women carrying male fetuses were detected by polymerase chain reaction.Results The PY3.4 position hybridization rates were 31.5%0 and 38% respectively in sorted GPA positive cells of 3 samples by FISHand the results of GPA+ cells of the 3 cases by PCR were positivenegative and positive respectivelywhich were manifested by the gender after aborticide.

选用红细胞特异单克隆抗体Glycophorin A结合流式细胞术成功地从3例妊娠16～24周的孕妇血中分离出NRBCs，并用Y-染色体特异重复序列DNA探针(PY3.4)荧光原位杂交及聚合酶链反应技术检测Y-染色体特异序列结果 FISH结果，3例分选出的GPA阳性细胞中PY3.4杂交阳性率分别是31.4%、0、38%；其PCR结果，分别为阳性、阴性、阳性，并在引产后得到性别证明。

The major reason maybe that the systemic errors in indirect hybridizing are more excessive than direct hybridizing.

结果表明，间接杂交的Log值的变化幅度要比直接杂交的大，主要原因是因为其实验系统误差要高于直接杂交

According to Griffing's method 1, diallel crossing was built and CA was analyzed. Variance analysis showed that there were the greatly significant difference among the both of GCA in inbreeds and SCA in crosses, respectively. The high GCA of 18-599 and 18-599 and the low SCA variance of their crosses showed the strong and stable heredity on the both characters. Taking them as parents, the generations with high and stable induction and regeneration will be gotten.

2选用在配合力、抗病性、农艺性状等玉米杂交育种性状与幼胚培养胚性愈伤组织诱导率和胚性愈伤组织绿苗发生数等转基因工程育种性状都表现相对十分优异的18-599、18-599和综31等3个自交系，以及杂交育种性状表现优良、但转基因工程育种性状表现极差的A318，P138和R15等3个自交系为亲本，按照Griffing方法1组配双列杂交，进行Griffing的配合力分析。

Total RNAs from KAx-3 cells and AK127 cells(developed for 14h) were isolated. After the reverse transcription and PCR reaction, two distinct differential fragments were acquired., fragment A was from KAx-3 cells and fragment C was from AK127 cells. After retriving and reamplifying the differentially expressed fragments, white-blue plaqueselection, the fragments were purified. Northern blot proved that fragment A was from KAx-3 cells and fragment C was from AK127 cells. The results of sequencing and researching for NCBI database have been showed: part sequence of fragment A shows 91% similarity to the gene encoding DhkA, 92% similarity to the gene encoding DhkF, 91% similarity to the gene encoding STATc, 97% similarity to the homoeobox gene encoding protein. These genes play important part in controlling cell differentiation and cell proportion in Dictyostelium discoideum.

本研究通过提取盘基网柄菌发育14小时的野生型KAx-3细胞和突变型AK127细胞的总RNA，运用mRNA差异显示法分离出了两条明显的差异表达片段，其中片段A来自野生型KAX-3细胞，片段C来自突变型Ak127细胞；并通过凝胶回收差异片段、对差异片段进行再次PCR、蓝白斑筛选克隆、提取质粒、酶切电泳纯化差异片段；接着进行Northern杂交的结果表明，片段A只与野生型KAx-3细胞的总RNA有杂交信号，片段C只与突变型AK127细胞的总RNA有杂交信号，这就排除了差异片段假阳性的可能；最后通过测序，搜索NCBI BLAST数据库发现：片段A的小部分序列与编码组氨酸激酶DhkA基因中一段序列的相似性高达91%，与编码组氨酸激酶DhkF基因中的一段序列相似性高达92%，与编码STATc蛋白基因的一段序列相似性达91%，以及与编码同源框蛋白的基因中的一段序列相似性达97%，这些基因在盘基网柄菌细胞分化和细胞比例调控过程中起着相当重要的作用，这些数据进一步说明了突变细胞不能完成发育的原因。

Oocyte samples from one group were collected to detect the presence and integration of HBV DNA within cells and chromosomes using PCR, Southern blot, dot hybridization and fluorescence in situ hybridization. The female animals from another group were mated with their normal males, respectively. Their zygotes, 2-cell embryos were collected to detect the integration of HBV DNA in the female pronuclei of zygotes and the replication and expression of HBV genes in the 2-cell embryos using FISH, RT-PCR and immunofluoresence assay.(1) PCR detected positive bands in the tested oocyte samples fromgoldon hamster and mice. Southern blot revealed clear hybridization signals in PCR products.

研究用金黄地鼠和小鼠建立实验动物模型：将卵巢内注射HBV DNA的实验动物分成两组，一组注射后进行超排卵，收集卵巢和输卵管的卵母细胞，用PCR、Southern杂交，斑点杂交和荧光原位杂交(fluorescence in situ hybridization、FISH)检测HBV在卵母细胞内的存在和染色体上的整合；另一组超排雌鼠与正常雄鼠合笼，收集受精卵和2-细胞胚，用FISH、RT-PCR和免疫荧光检测技术分别研究HBV基因在受精卵雌原核上的整合以及在2-细胞胚中的复制与表达。

As a result,1A360 probe did hybridize strongly to Tester DNA sample,but weakly to Driver DNA.Thus,our two rounds of RDA are proved to be specific andeffective.Because of not hybridizing to the liver tissue DNA of rat and the thymicDNA of bull,so the probe is species-specific and gained from human chromosome.

该探针不能与鼠肝组织和小牛胸腺组织DNA杂交，说明它来源于人源染色体并有种属的特异性。1A360基因片段探针还与多种胃癌细胞系，胃癌组织DNA有较强的杂交信号，与其它肿瘤DNA有强弱不等的杂交信号。

Major difference of gene expression occurred in the qualitative level, especially cross-fertilized kernel specifically expressed pattern.

对三个不同优势的小麦杂交种子和自交种子及其正反杂交种与其双亲种子发育前期基因表达研究表明，杂交种子和自交种子之间存在明显的基因表达差异，且以质的特异表达为主；不同优势的正反杂交当代种子与亲本间基因表达发生了显著的改变，进一步分析发现单亲表达与杂种优势存在一定的正向关系，而杂种沉默表达与杂种优势存在一定的负向关系。

In order to understand the molecular basis of heterosis, modified differential display of mRNA was used to characterize the difference in gene expression between cross-fertilized and self-fertilized kernels, reciprocal cross-fertilized kernels and their parents during the early stages of seed development by using 3 wheat hybrids with different level of heterosis.

本研究旨在利用mRNA差异显示技术探讨小麦杂交种子和自交种子以及正反杂交种子与其双亲在种子发育前期的基因表达差异，同时对杂种优势利用的模式作物玉米的杂交种子和自交种子的基因表达差异也进行了研究，并对典型的差异表达片段进行了克隆测序，为揭示杂种优势形成的分子机理积累了更多的资料，取得的主要结果如下

This paper study on the hybridization of different strain soft-shelled turtle by traditioncrossbreeding. Then compare the difference of body character, components of muscle andamino acid and the fatty acid composition, The conclusion is as fellows.

本研究采用杂交育种技术，对不同地方品系中华鳖进行杂交，对照种鳖及杂交子代鳖的体表形态特征；分析肌肉成分，肌肉和裙边中氨基酸和脂肪酸的差异；同时本文还进行了LPS对中华鳖免疫增强效果的研究，主要结果如下： 1。

In the United States, chronic alcoholism and hepatitis C are the most common ones.

在美国，慢性酒精中毒，肝炎是最常见的。

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