The commercial and agricultural applications of animal transgenesis

The potential for commercial application of transgenic technologies in domestic animals is discussed in relation to the areas where a significant impact on agriculture might be expected. These are the endocrine system, novel biochemical pathways, structural proteins of milk and of textile fibers, and the immune system. Manipulation of the endocrine system has been investigated for some years and it is clear that very accurate control is needed over gene expression if this approach is to prove commercially useful. The area most advanced in commercial application is the production of high-value pharmaceutical proteins in the mammary glands of domestic animals. Other applications that are discussed remain to be proven in larger animals despite being demonstrated laboratory test animals. These include a functional cysteine biosynthetic pathway and a functional glyoxylate cycle transferred from bacteria to mice, and alterations to the proteins of hair that change the physical properties of the resultant fibers. Research is also actively directed toward novel approaches for providing domestic animals with resistance to insects.     

羊输卵管液中发情相关糖蛋白的性质研究

哺乳动物输卵管液为受精和早期受精卵的发育提供了一个理想的生理生化环境。人们对输卵管液的成分已进行了初步研究,其中发情相关糖蛋白（ＥＧＰ）是被研究的成分之一。ＥＧＰ仅存在在排卵核受精前后的输卵管液中。当受精卵进入子宫区,输卵管液中的ＥＧＰ就消失了。显然,ＥＧＰ是与哺乳动物的早期发育过程密切相关的。尽管人们对ＥＧＰ的确切生理功能尚不清楚,但作为第一步,首先弄清ＥＧＰ的理化性质是十分重要的。本文目的在于研究羊输卵管液中ＥＧＰ的理化性质。本文采用单向（１Ｄ）、双向（２Ｄ）ＳＤＳ－聚丙烯胺凝胶电泳（ＰＡＧＥ）和等电聚胶电泳（ＩＥＦ）,结合Ｗｃｓｔｅｒｎｂｌｏｔ技术对ＥＧＰ的性质进行了研究。由于单克隆抗体及花生凝集素（ＰｅａｎｕｔＡｇｇｌｕｔｉｎｉｎ）和ＥＧＰ相结合的专一性很高,所以不必事先对ＥＧＰ进行纯化就可直接进行分析,我们的结果证明羊ＥＧＰ包括两种蛋白质,一种是酸性蛋白,它的等电点为数４．５;另一种是碱性蛋白,其等电点是８．０;这两种蛋白亚基的分子量相同,都是１０ＫＤ。从不同物种间ＥＧＰ这些理化数据上比较,羊和狒狒的非常接近,但和其他动物的有很多不同。我们认为来自不同动物的ＥＧＰ可能是一类性质相同的蛋白质,具有相     

Linkage mapping of melanoma (MLM) using 172 microsatellite markers

The incidence of malignant melanoma is currently increasing faster than any other cancer and in 5-12% of cases occurs in a familial context in which the disease cosegregates as an autosomal dominant trait. To identify the location of genes that predipose individuals to familial melanoma (MLM), we have carried out linkage analysis in three large Australian melanoma pedigrees using 172 microsatellite markers spread across all autosomes. Three additional smaller families were typed for 70 of the same markers. In five of the six families we found lod scores between 1.0 and 2.3, which may provide evidence for the location of melanoma genes in proximity to some of these markers. If this turns out to be the case, these data potentially demonstrate that MLM is genetically heterogeneous since there was no marker for which all families gave significantly high LODs. These data provide the foundation for an exclusion map for melanoma and, more importantly, high-light areas of the genome for others to substantiate the potential positions of some of the genes that may be responsible for susceptibility to MLM.     

Evaluation of a cosmid contig physical map of human chromosome 16.

A cosmid contig physical map of human chromosome 16 has been developed by repetitive sequence finger-printing of approximately 4000 cosmid clones obtained from a chromosome 16-specific cosmid library. The arrangement of clones in contigs is determined by (1) estimating cosmid length and determining the likelihoods for all possible pairwise clone overlaps, using the fingerprint data, and (2) using an optimization technique to fit contig maps to these estimates. Two important questions concerning this contig map are how much of chromosome 16 is covered and how accurate are the assembled contigs. Both questions can be addressed by hybridization of single-copy sequence probes to gridded arrays of the cosmids. All of the fingerprinted clones have been arrayed on nylon membranes so that any region of interest can be identified by hybridization. The hybridization experiments indicate that approximately 84% of the euchromatic arms of chromosome 16 are covered by contigs and singleton cosmids. Both grid hybridization (26 contigs) and pulsed-field gel electrophoresis experiments (11 contigs) confirmed the assembled contigs, indicating that false positive overlaps occur infrequently in the present map. Furthermore, regional localization of 93 contigs and singleton cosmids to a somatic cell hybrid mapping panel indicates that there is no bias in the coverage of the euchromatic arms.     

Effect of anti-oestradiol-17B antibodies on the reproductive response of ewes superovulated with PMSG

A field experiment was conducted to examine the effect of anti-oestradiol-17B antibody titre on the oestrous and ovulatory responses of ewes to low (600 i.u.) or high (1200 i.u.) doses of pregnant mare's serum gonadotrophin (PMSG). Merino ewes were treated with intravaginal sponges and were subsequently used as vehicle-treated controls or were immunized to produce reciprocal anti-oestradiol-17B antibody titres less than 1000 or greater than 1000. Ewes were then treated with PMSG and the incidence of oestrus and ovulation, ovulation rate, and yield of embryos recorded. Treatment of immune ewes with 1200 i.u. PMSG resulted in both a higher proportion of ewes ovulating and a higher ovulation rate than in immune ewes treated with 600 i.u. (86% v. 67% and 13.4 v. 6.0 respectively). As anti-oestradiol-17B titres increased there was a reduction in the proportion of ewes exhibiting oestrus. The proportion of ewes ovulating decreased as antibody increased in ewes treated with 600 i.u. PMSG but not in those treated with 1200 i.u., suggesting an increased positive feedback of oestradiol with high PMSG doses. Fertilization rates were highest at the lower PMSG dose (68% v. 42%) and increased with increasing titre. Overall, there was no increase in ovulation rate or in yield of embryos over control values from either low (less than 1000) or high (greater than 1000) antibody titres.     

Characterization of the Melanoma miRNAome by Deep Sequencing

Background

MicroRNAs (miRNAs) are 18–23 nucleotide non-coding RNAs that regulate gene expression in a sequence specific manner. Little is known about the repertoire and function of miRNAs in melanoma or the melanocytic lineage. We therefore undertook a comprehensive analysis of the miRNAome in a diverse range of pigment cells including: melanoblasts, melanocytes, congenital nevocytes, acral, mucosal, cutaneous and uveal melanoma cells.

Methodology/Principal Findings

We sequenced 12 small RNA libraries using Illumina''s Genome Analyzer II platform. This massively parallel sequencing approach of a diverse set of melanoma and pigment cell libraries revealed a total of 539 known mature and mature-star sequences, along with the prediction of 279 novel miRNA candidates, of which 109 were common to 2 or more libraries and 3 were present in all libraries.

Conclusions/Significance

Some of the novel candidate miRNAs may be specific to the melanocytic lineage and as such could be used as biomarkers to assist in the early detection of distant metastases by measuring the circulating levels in blood. Follow up studies of the functional roles of these pigment cell miRNAs and the identification of the targets should shed further light on the development and progression of melanoma.