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91.
92.
小麦×玉米杂交后代的蛋白质及酯酶同工酶分析 总被引:9,自引:0,他引:9
以8 个普通小麦(Triticum aestivum L.)品种为母本,2 个栽培玉米(Zea m ays L.)品种为父本杂交所获得的F2 代在形态上出现了明显变异。对其籽粒进行蛋白质电泳分析,得到了如下主要结果:杂交后代的蛋白质谱带较母本有了很大的变异,主要集中在高分子量麦谷蛋白(HMW-Glu)区域。杂交后代的蛋白质谱带由5 种类型构成:1.母本型,占全部测试籽粒的22.6% ;2.附加型,占14.3% ;3.互补型,占15.5% ;4.杂种型,占30.9% ;5.缺失型,占16.7% 。对“矮杆早”ד紫粘”的F2 代籽粒进行酯酶同工酶电泳分析发现,变异主要发生在EST-1 区。由此看来,小麦×玉米的杂合子中玉米染色体在被排除前后,可以诱发小麦染色体组发生遗传变异 相似文献
93.
John O. Hui John Le Viswanatham Katta Robert Rosenfeld Michael F. Rohde Mitsuru Haniu 《Journal of Protein Chemistry》1996,15(4):351-358
Human neurotrophin-3 (NT-3) is a member of the nerve growth factor (NGF) family of neurotrophic factors, and the recombinant protein is being developed as a therapeutic for neurodegenerative diseases. The final product purity and lot-to-lot variation are monitored routinely by peptide mapping. However, only the N-terminal region of NT-3 was susceptible to proteolysis under native conditions. Complete digestion required that the protein be chemically modified by reduction and S-alkylation prior to proteolysis. Complete proteolytic degradation of the protein was achieved simply by an intial denaturation of NT-3 in 6 M guanidinium chloride (pH 6) for 2 hr at 37°C, followed by a tenfold dilution with the digestion buffer (0.1 M Tris-HCl, 1 mM CaCl2 at pH 7.0) and immediate addition of chymotrypsin at 1% by weight. Direct comparison of the peptide map with an identical aliquot that had been reduced and alkylated also allowed the establishment of the cystine linkages present in NT-3: Cys14 to Cys79, Cys57 to Cys108, and Cys67 to Cys110. This disulfide structure is homologous to the NGF family of neurotrophic factors. 相似文献
94.
New telomere formation coupled with site-specific chromosome breakage in Tetrahymena thermophila. 总被引:5,自引:2,他引:3
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Programmed chromosome breakage occurs in many ciliated protozoa and is accompanied by efficient new telomere formation. In this study, we have investigated the relationship between programmed chromosome breakage and telomere formation in Tetrahymena thermophila. Using specially constructed DNA clones containing the breakage signal Cbs in transformation studies, we have determined the locations of telomere addition around the breakage sites. They occur at variable positions, over 90% of which are within a small region (less than 30 bp) starting 4 bp from Cbs. This distribution is independent of the nucleotide sequence in the region or of the orientation of Cbs. In five of six cases determined, these sites occur at or before a T, and in the remaining case, the site occurs at or before a G. When sequences devoid of G or T are placed in this region, telomere addition still occurs within the region to maintain a similar distance relationship with Cbs. This efficient and healing process appears to be associated specifically with Cbs-directed breakage, since it does not occur when DNA ends are generated by restriction enzyme digestion. These results suggest a strong mechanistic link between chromosome breakage and telomere formation. 相似文献
95.
Sequence Variations in the Bovine Growth Hormone Gene Characterized by Single-Strand Conformation Polymorphism (Sscp) Analysis and Their Association with Milk Production Traits in Holsteins 总被引:5,自引:0,他引:5
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Sequence variations in the bovine growth hormone (GH) gene were investigated by single strand conformation polymorphism (SSCP) analysis of seven amplified fragments covering almost the entire gene (2.7 kb). SSCPs were detected in four of these fragments and a total of six polymorphisms were found in a sample of 128 Holstein bulls. Two polymorphisms, a T->C transition in the third intron (designated GH4.1) and an A->C transversion in the fifth exon (designated GH6.2), were shown to be associated with milk production traits. GH4.1(c)/GH4.1(c) bulls had higher milk yield than GH4.1(c)/GH4.1(t) (P <= 0.005) and GH4.1(t)/GH4.1(t) (P <= 0.0022) bulls. GH4.1(c)/GH4.1(c) bulls had higher kg fat (P <= 0.0076) and protein (P <= 0.0018) than GH4.1(c)/GH4.1(t) bulls. Similar effects on milk production traits with the GH6.2 polymorphism were observed with the GH6.2(a) allele being the favorable allele. The average effects of the gene substitution for GH4.1 and GH6.2 are similar, with +/-300 kg for milk yield, +/-8 kg for fat content and +/-7 kg for protein content per lactation. The positive association of GH4.1(c) and GH6.2(a) with milk production traits may be useful for improving milk performance in dairy cattle. 相似文献
96.
Evolutionary Conservation of Sequences Directing Chromosome Breakage and Rdna Palindrome Formation in Tetrahymenine Ciliates 总被引:2,自引:0,他引:2
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Extensive, programmed chromosome breakage occurs during formation of the somatic macronucleus of ciliated protozoa. The cis-acting signal directing breakage has been most rigorously defined in Tetrahymena thermophila, where it consists of a 15-bp DNA sequence known as Cbs, for chromosome breakage sequence. We have identified sequences identical or nearly identical to the T. thermophila Cbs at sites of breakage flanking the germline micronuclear rDNA locus of six additional species of Tetrahymena as well as members of two related genera. Other general features of the breakage site are also conserved, but surprisingly, the orientation and number of copies of Cbs are not always conserved, suggesting the occurrence of germline rearrangement events over evolutionary time. At one end of the T. thermophila micronuclear rDNA locus, a pair of short inverted repeats adjacent to Cbs directs the formation of a giant palindromic molecule. We have examined the corresponding sequences from two other Tetrahymena species. We find the sequence to be partially conserved, as previously implied from analysis of macronuclear rDNA, but of variable length and organization. 相似文献
97.
二核苷酸重复多态性的非同位素检测及其在基因诊断中的应用 总被引:9,自引:1,他引:8
本文报道了一种检测二核苷酸重复多态性的简便的非同位素法,利用重复序列两侧的特异引物进行PCR扩增,产生的等位片段在薄层变性聚丙烯酰胺凝胶电泳上分离,再用灵敏的银染法显色。该方法不需要标记PCR产物,简便、快速,分辨率可达1bp,并可用多对引物同时进行多重PCR分析。用此方法对DMD家系成员dystrophin基因的5'-脑型外显子止游区和3'-非翻译区的两个(CA)。位点进行了扩增片段长度多态性分 相似文献
98.
利用PCR方法对单纯疱疹病毒Ⅱ型糖蛋白D(HSV-2gD)基因进行了修饰,在其5'端删去约500bp的非编码区,仅保留ATG上游7个bp。将修饰后的HSV-2gD基因插入到带有痘苗病毒天坛株TK基因区段的痘苗表达质粒pJSA1175,置于痘苗病毒P7.5k早/晚期启动子控制下。将此重组质粒用脂质体Lipofectin方法转染已受野型TK ̄+痘苗病毒天坛株感染的TK ̄-143细胞,通过同源重组机制和标志基因LacZ产物的蓝斑显色作用,以及BudR试剂对TK表型的选择压力,筛选出整合有HSV-2gD基因的重组痘苗病毒。Southem杂交表明,HSV-2gD基因已正确地插入痘苗病毒TK基因区内;间接免疫荧光检测显示,HSV-2gD蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗HSV-2gD中和抗体。用重组病毒免疫小鼠,3周后可使94%(17/18)的小鼠对抗HSV-2的致死量攻击,表明重组病毒具有明显的免疫保护作用。 相似文献
99.
100.
用人重组肿瘤坏死因子-α(Tumornecrosisfactor-α,TNF-α)和人天然α干扰素(Interferon-α-,IFN-α)在人胚胎肺纤维母细胞(HEF)和Hep-2细胞系上对常见呼吸道病毒所致细胞病变抑制进行比较观察。病毒包括不同型别的腺病毒5株,单疱病毒Ⅰ型(HSV-I)1株,鼻病毒1株,仙台病毒1株,VSV1株。结果提示TNF-α和IFN-α均具有广谱抗病毒活性。TNF-α的抑毒作用能被TNF-α申抗和IFN-β单抗完全去除,被IFN-α单抗部分去除TNF-α的抗病毒效应。TNF--α中和试验的结果提示:TNF抗病毒活性仍为IFN-β诱生所介导。 相似文献