An essential component of functional genomics studies is the sequence of DNA expressed in tissues of interest. To provide a resource of bovine-specific expressed sequence data and facilitate this powerful approach in cattle research, four normalized cDNA libraries were produced and arrayed for high-throughput sequencing. The libraries were made with RNA pooled from multiple tissues to increase efficiency of normalization and maximize the number of independent genes for which sequence data were obtained. Target tissues included those with highest likelihood to have impact on production parameters of animal health, growth, reproductive efficiency, and carcass merit. Success of normalization and inter- and intralibrary redundancy were assessed by collecting 6000–23,000 sequences from each of the libraries (68,520 total sequences deposited in GenBank). Sequence comparison and assembly of these sequences was performed in combination with 56,500 other bovine EST sequences present in the GenBank dbEST database to construct a cattle Gene Index (available from The Institute for Genomic Research at http://www.tigr.org/tdb/tgi.shtml ). The 124,381 bovine ESTs present in GenBank at the time of the analysis form 16,740 assemblies that are listed and annotated on the Web site. Analysis of individual library sequence data indicates that the pooled-tissue approach was highly effective in preparing libraries for efficient deep sequencing.
Abstract We report the most extensive physically anchored linkage map for cattle produced to date. Three-hundred thirteen genetic markers ordered in 30 linkage groups, anchored to 24 autosomal chromosomes (n = 29), the X and Y chromosomes, four unanchored syntenic groups and two unassigned linkage groups spanning 2464 cM of the bovine genome are summarized. The map also assigns 19 type I loci to specific chromosomes and/or syntenic groups and four cosmid clones containing informative microsatellites to chromosomes 13, 25 and 29 anchoring syntenic groups U11, U7 and U8, respectively. This map provides the skeletal framework prerequisite to development of a comprehensive genetic map for cattle and analysis of economic trait loci (ETL).
The murine E‐protein gene ME1 encodes a non‐tissue‐specific, helix‐loop‐helix transcription factor that is associated with morphological development. ME1 gene expression is regulated by a TATA‐less promoter that contains multiple Sp1 consensus elements, E‐boxes, and a novel transcription initiation site. In this study, we compared DNA homologous to the ME1 promoter from vertebrate species ranging from frog to human. A region of striking sequence similarity was identified in a region corresponding to the ME1 transcription initiation site (ME1 Inr). Within this region, a poly d(A) tract and a 9‐bp inverted repeat (5′‐GTCCGCCTG) were highly conserved in all species that were examined. Protein complexes that recognized these DNA elements were present among distant vertebrates (frog, chick, monkey and human), and were able to bend the ME1 Inr to a similar extent (∼60°) as the previously described murine MBPα and MBPβ proteins. Collectively, these results suggest that an ME1 Inr‐like element and its associated proteins functioned in an ancestral vertebrate more than 350 million years ago.
Human choriogonadotropin (hCG) is a heterodimeric glycoprotein hormone. The alpha subunit comprises 92 amino acids, of which 6 are Lys residues (Morgan, F.G., Birken, S., and Canfield, R.E. (1975) J. Biol. Chem. 250, 5247-5258). Our photoaffinity-labeling studies indicate that several of these Lys residues make contact with the lutropin receptor and are covalently cross-linked to the receptor. Lys-91 of the alpha subunit is of interest because deletion of the two alpha C-terminal residues, Lys-91 and Ser-92, results in a significant reduction in the bioactivity of lutropin and thyrotropin (Cheng, K.-W., Glazer, A.N., and Pierce, J.G. (1973) J. Biol. Chem. 248, 7930-7937). To determine the importance of Lys-alpha 91, we substituted it with Arg, Met, or Glu. The resulting mutant alpha cDNA constructs were co-transfected into CHO cells with the wild type hCG beta cDNA construct. Secreted hCG dimers were assayed for binding to receptors on porcine granulosa cells and stimulation of cAMP synthesis in a murine Leydig tumor cell line. The natural hCG, wild type hCG, and all mutant hCGs recognized the receptor, although with somewhat divergent affinities. However, there was a striking difference in the ability of cAMP induction. The natural hCG, wild type hCG, and Lys-91—-Arg mutant hCG induced cAMP synthesis, whereas the Lys-91—-Met and Lys-91—-Glu mutants did not. These results demonstrate that Lys-91 is important for receptor modulation in the stimulation of cAMP synthesis.
We have cloned a minisatellite tandem array (XTA) from Xenopus laevis that contains approximately 200 copies of the 20-bp repeat 5′-CCAACAGCCTGCCCATCCAT-3′. The XTA sequence is present only once per haploid genome and is polymorphic with respect to repeat number and location of flanking restriction endonuclease sites. Although the 20-bp repeat has not previously been described, flanking sequences suggest that it lies proximal to coding regions in the Xenopus genome. Key words : repetitive DNA, minisatellite DNA, VNTR, evolution, tetraploid, polymorphic.
본 연구에서는 천연 암반심해수를 가축 음용수로 개발하고자 2.3% 염분을 함유하고, 칼륨(K), 마그네슘(Mg), 칼슘(Ca), 나트륨(Na), 철(Fe), 망간(Mn), 아연(Zn), 구리(Cu) 등의 광물질을 함유한 암반심해수를 0.5% 및 1%로 희석하여 흰쥐에 공급하며 면역 반응과 항산화 활성에 미치는 영향을 조사하였다. 공시동물로 Sprague Dawley종의 수컷 흰쥐 24마리를 6마리 씩 대조군과 3개의 처리군으로 임의 배치하고, 대조군은 정제수를 급여하였고 처리군은 각각 0.5% 암반심해수, 1% 암반심해수, 지장수를 공급하여 4주 동안 사육하였다. 실험 결과, 1%의 암반수의 경우가 증체량 및 사료섭취량, 사료효율이 우수한 것으로 나타났다. 총음수량은 대조군 보다 0.5% 암반심해수의 경우는 49%, 1% 암반심해수는 22.8%, 지장수는 40.5% 가량 더 많은 것으로 나타났다(p<0.05). 각 조직무게의 경우, 지장수군의 신장주위지방과 정소상체지방은 각각 32%, 25% 적은 것으로 나타났다(p<0.05). B 세포와 T 세포 자극능은 처리군중에서 0.5% 암반심해수 급여군이 가장 높아 대조군보다 44.7%와 207% 증가한 것으로 나타났다(p<0.05). 또 0.5% 암반심해수가 혈청 내 triglyceride는 4%, Free fatty acid는 4%, LDL- cholesterol은 45.9% 감소시키는 등 지질성분 변화에 가장 긍정적인 효과를 나타내었다. 항산화 활성의 경우 TBARS의 양은 0.5% 암반심해수는 21.8%, 1% 암반심해수는 34.7%, 지장수는 20.8% 정도 감소하였다. catalase 활성은 0.5% 암반심해수가 약 2.9배, 지장수가 약 1.6배 증가하였으며, Cu/Zn SOD 활성은 0.5% 암반심해수,가 약 1.5배, 1% 암반심해수가 약 1.7배 ,지장수가 약 1.2배 증가하였으며, MnSOD 활성은 모든 처리군이 대조군에 비해 약간 증가하였다. 이상의 결과로부터 가축에 0.5% 암반심해수의 급여는 LDL 콜레스테롤의 농도가 감소되고, 면역 활성 및 항산화효소 활성이 높아지는 등 고급 축산물생산의 음용수로 유익할 것으로 고찰되었다. This experiment was conducted to study the effects of the natural deep sea water, which contained approximately 2.3% salt, and various minerals of K, Mg, Ca, Na, Fe, Mn, Zn, Cu etc, on the immune response and antioxidant activity in rats. 24 Sprague Dawley rats were randomly allotted to a control group and 3 treatment groups. Control rats were supplied with filtered tap water, and each treatment group rats were supplied with 0.5% deep sea water, 1% deep sea water and Jijangsoo, respectively, which is upper clear water separated from sediment by the clay. Feed and water were provided ad libitum throughout the experiment that lasted for 4 weeks. The results showed that 1% deep sea water group showed the highest values in weight gain, feed intake, and feed efficiency than those of other groups. The levels of water intake of 1%- and 0.5%-deep sea water, and Jijangsoo group were 49.1%, 22.8%, and 40.5% higher than that of control group, respectively. The Jijangsoo group rats showed that perirenal and epididymal adipose tissue weights were decreased by 32% and 25%(p<0.05), respectively, when compared to control group rats. There were no remarkable differences of serum glucose concentration among all experimental groups. However, insulin concentration of experimental groups were remarkably increased in order of Jijangsoo (4.54), 1% deep sea water (3.70), 0.5% deep sea water (3.25)(p<0.05). B cell and T cell stimulation were increased about 44.7% and 207%, respectively, by 0.5% deep sea water in comparison with control (p<0.05). TBARS values of 0.5 % deep sea water group were significantly lower than that of control(p<0.05). Catalase and SOD activities of 0.5 % deep sea water group were 200% and 47% higher than that of control, respectively. From the results, it can be concluded that the supply of natural deep sea water can slightly improve the physiological activity which modulates immune response and antioxidant activity in rats.
Human choriogonadotropin (hCG) and follitropin (FSH) belong to the glycoprotein hormone family. These hormones are heterodimers and composed of a common alpha subunit and a distinct beta subunit which confers receptor-binding specificities. In addition to this structural similarity, they share a similar signal pathway involving G protein, adenylyl-cyclase and induction of cAMP synthesis. Therefore, a presumptive relationship of these common structure and function has been the subject of extensive past investigations, but a definitive clue has been elusive. As a step to address this important issue, a series of recombinant mutants of hCG and human FSH were generated in which the COOH-terminal amino acids of the alpha subunit were successively removed or substituted. Furthermore, a set of peptides were synthesized with sequences corresponding to different regions of the alpha subunit. Deletion of the alpha COOH-terminal Ser92 had no effect on receptor-binding or cAMP induction by FSH and hCG. Truncation of alpha Lys91-Ser92 or alpha His90-Lys91-Ser92 abolished the ability of both hormones to induce cAMP synthesis. It significantly reduced receptor binding of FSH but not hCG. The different functions of the alpha COOH-terminal region are further noticed with a peptide corresponding to the last 10 amino acids of alpha. It failed to bind to the FSH receptor but was capable of binding to the LH/CG receptor and stimulating cAMP synthesis. These results are the first conclusive evidence that alpha His90-Lys91 play an essential role in cAMP induction of both hormones. In contrast to this common role, they are necessary for FSH binding to the FSH receptor but not for hCG binding to the LH/CG receptor. The hCG alpha COOH-terminal region makes direct contact with the LH/CG receptor, and this low affinity contact is necessary and sufficient to activate the receptor for signal generation. This conclusion is supported by the study using mutant hCGs in which either alpha His90 or Lys91 was substituted.
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