New telomere formation coupled with site-specific chromosome breakage in Tetrahymena thermophila.

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.     

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

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.     

Evolutionary Conservation of Sequences Directing Chromosome Breakage and Rdna Palindrome Formation in Tetrahymenine Ciliates

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.     

籼稻232蜡质基因转录起始位点的鉴定

Ｎｏｒｔｈｅｒｎｂｌｏｔ杂交分析和蜡质基因ｃＤＮＡ的序列分析表明水稻蜡质基因的转录本可能延伸到翻译起始密码子（ＡＴＧ）上游１２ｋｂ处。据此设计了２１Ｎｔ的寡核苷酸引物,并以籼稻２３２胚乳ＲＮＡ为模板,以引物延伸法确定籼稻２３２蜡质基因的转录起始点,籼稻蜡质基因的转录起始旁邻顺序ＣＴＣＡＣＣＡ与高等植物基因的转录起始点一致顺序ＣＴＣＡＴＣＡ仅相差１个碱基。通过顺序比较,对东乡野生稻蜡质基因中的转录起始位点的位置,以及对此两稻种中ＴＡＴＡ盒的可能顺序进行了讨论。     

肿瘤坏死因子－α（TNF－α）对常见呼吸道病毒的抑制作用

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。     

旋扭山绿豆根瘤菌MXDI6菌株氢酶诱导表达

在自生异养条件下,旋扭山绿豆根瘤菌ＭＸＤＩ６菌株的氢酶诱导表达受气相、ｐＨ值、镍等因子影响：氢酶表达的最适氧浓度为４％,最适氢浓度为１５％,二氧化碳没有明显影响;氢酶表达的ｐＨ值以５．０—６．０为宜;０．５μｍｏｌ／ＬＮｉＣｌ２明显促进吸氢活性,但镍浓度大于１μｍｏｌ／Ｌ则抑制吸氢活性．     

Ultrastructure of the spermatozoa and eggs of the ocean pout (Macrozoarces americanus L.), an internally fertilizing marine fish

Ultrastructure of sperm and eggs of the ocean pout (Macrozoarces americanus L.), an internally fertilizing marine teleost, was examined by scanning and transmission electron microscopy. The results showed that the sperm do not have an acrosome but have a very long mid-piece (one to two times the sperm head length) containing numerous well-developed elongated mitochondria. The sperm also have two tails (is biflagellate) each consisting of nine peripheral and one central pair (9 ± 2) of microtubules. This long mid-piece and the biflagellate nature of the sperm appear to be associated with the long life-span of the sperm and with sperm dispersal in the ovary to fertilize the eggs internally. The ocean pout eggs are enveloped by a porous chorionic membrane similar to that found in other teleosts but have two micropyles, a condition likely related to a mechanism of egg fertilization which increases the egg fertlity in the presence of low sperm numbers. Following insemination, some biochemically undefined excretions appeared on the surface of fertilized eggs and led to the acquisition of adherent capability of the eggs which formed a tightly associated egg mass in sea water. © 1995 wiley-Liss, Inc.     

An rRNA variable region has an evolutionarily conserved essential role despite sequence divergence.

Regions extremely variable in size and sequence occur at conserved locations in eukaryotic rRNAs. The functional importance of one such region was determined by gene reconstruction and replacement in Tetrahymena thermophila. Deletion of the D8 region of the large-subunit rRNA inactivates T. thermophila rRNA genes (rDNA): transformants containing only this type of rDNA are unable to grow. Replacement with an unrelated sequence of similar size or a variable region from a different position in the rRNA also inactivated the rDNA. Mutant rRNAs resulting from such constructs were present only in precursor forms, suggesting that these rRNAs are deficient in either processing or stabilization of the mature form. Replacement with D8 regions from three other organisms restored function, even though the sequences are very different. Thus, these D8 regions share an essential functional feature that is not reflected in their primary sequences. Similar tertiary structures may be the quality these sequences share that allows them to function interchangeably.