A novel human gene spindlin1, encoding a protein localized in the cell nucleus and inducing NIH3T3 cell''s transformation

A novel human gene, spindlin1, recently cloned in our laboratory, is highly expressed in the tissue of ovary cancer. To study its biological function, a vector expressing green fluorescent-spindlin1 fusion protein was constructed and transfected into COS-7 and NIH3T3 cells by lipofectamine methods. The results showed that the fusion protein pEGFP-N1-spindlin1 was localized in the nucleus of COS-7 and NIH3T3 cells. NIH3T3 cells which could stably express spindlin1 as a result of RT-PCR analysis compared with the parental NIH3T3 cells displayed a complete morphological change, improved the cell growth and increased the percentage of cells in G2/M phase (12.6% vs control cells at 3.4%). Furthermore, overexpressed spindlin1 cells formed colonies in soft agar, more motile in migration assay in vitro and formed tumors in nude mice. Our findings provide direct evidence that spindlin1 gene may be a prooncogene which is associated with tumorigenesis.     

人神经节苷脂诱导分化相关蛋白cDNA的克隆、染色体定位和初步功能研究

通过构建,筛选人18周胎脑cDNA文库,克隆到一条与神经节苷脂诱导分化相关蛋白高度同源的新基因,经HUGO／GDB人类基因命名委员会的同意命名为GDAP1L1,进行新基因的全序列测定,RH定位分析,Blast分析及生物学信息分析,Northern杂交提示GDAP1L1基因在胎脑中高度表达,但在成人脑组织中低表达,新的神经节甙脂诱导分化相关蛋白的表达和功能研究初步提示：全长新神经节苷脂诱导分化相关基因核苷酸序列长1163bp,RH定位分析新基因定位在染色体20q12区,BLASTN,BLASTP,TBLASTN分析新基因的蛋白质序列与人和鼠“神经节甘脂诱导分化相关蛋白1”有58％的同源性,而与人的另一条“类似神经节苷脂诱导分化相关蛋白1”的部分蛋白序列（47－253aa)的同源性达100％,新基因蛋白在3,5端分别多出46aa和114aa的长度,生物学住处分析证实神经节苷脂诱导分化相关基因与神经营养与细胞凋亡有密切关系,全长新神经节苷脂诱导分化相关基因是一个神经营养与发育及细胞周期调控,信号传导有关的基因,可能在肿瘤的发生中具有重要作用,其功能的进一步研究将为肿瘤机理的阐明提供思路。     

Expression of recombinant human lysozyme in the milk of transgenic mice

Human lysozyme is a 130-aa (amino acid) alkaline polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal of improving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomic DNA sequence of the human lysozme gene has been generated for the first time. From 83 mice generated by microinjection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene in lines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic mice was 0.2mg/mL. The antibacterial activity of the whey for transgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse was very low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides a strong basis for further studies into the possible application of rHlys express in mammary gland.     

Characterization of the human factor VIII gene

The complete 186,000 base-pair (bp) human factor VIII gene has been isolated and consists of 26 exons ranging in size from 69 to 3,106 bp and introns as large as 32.4 kilobases (kb). Nine kb of mRNA and protein-coding DNA has been sequenced and the mRNA termini have been mapped. The relationship between internal duplications in factor VIII and evolution of the gene is discussed.     

Isolation and cloning of a novel cDNA LDBl encoding human LIM domain binding protein

Primers for screening cDNA library have been designed according to EST AA453734 which is corresponding to the mouse LIM domain binding protein Ldb1. Arrayed human fetal brain cDNA library has been screened by PCR and routine hybridization method. A 2398 bp-cD-NA clone has been obtained. The cDNA encodes a 347 amino acids protein highly homologous to the mouse Ldb1, Xenopus Xldb1 and Drosophila Chip. It also contains an LIM binding domain and a nuclear localization signal. It has been named LDB1 (LIM domain binding protein 1), GenBank accession number is AF052389. Northern blot showed a 2.4 kb band, and the expression amounts of LDB1 in heart, brain and lung were considerably higher than those in other tissues.     

Comparative chromosome mapping of a conserved homoeo box region in mouse and human

Specific genes are assumed to regulate pattern formation in the mammalian embryo, but as yet none has been identified unequivocally. It is possible that such genes in mammals may be identified by virtue of a conserved coding sequence, because many of the Drosophila melanogaster homoeotic and segmentation genes, which have crucial roles in the regulation of segmental pattern formation during embryonic development, contain a 180-base pair (bp) DNA sequence, the homoeo box, and that sequences homologous to the Drosophila homoeo box are also present in 6-10 copies in higher animals, including mammals. Although the assumption that the homoeo box identifies genes responsible for pattern formation in mammals remains to be validated, it is a particularly attractive hypothesis given the strong conservation of homoeo boxes over vast evolutionary distances. Here we report the localization of a human homoeo box region, previously cloned and shown to contain two homoeo boxes within a sequence of 5-kilobases (kb), to the long arm of chromosome 17. We show that two single-copy homoeo box-flanking probes derived from this region strongly hybridize to single-copy restriction fragments in mouse genomic DNA and that these conserved homoeo box-flanking sequences map to mouse chromosome 11. This may be significant as several genes that map to chromosome 17 in human also map to chromosome 11 in the mouse, implying that a segment of mouse chromosome 11 is homologous to a region of human chromosome 17. Taken together, these data suggest that the homoeo box region detected with our probes is highly conserved in human and mouse.     

Isolation and cloning of a novel cDNALDB1 encoding human LIM domain binding protein

Primers for screening cDNA library have been designed according to EST AA453734 which is corresponding to the mouse LIM domain binding protein Ldbl. Arrayed human fetal brain cDNA library has been screened by PCR and routine hybridization method. A 2398 bp-cD-NA clone has been obtained. The cDNA encodes a 347 amino acids protein highly homologous to the mouse Ldbl,Xenopus Xldbl andDrosophila Chip. It also contains an LIM binding domain and a nuclear localization signal. It has been namedLDB1 ( LIM domain binding protein 1), GenBank accession number is AF052389. Northern blot showed a 2.4 kb band, and the expression amounts ofLDBI in heart, brain and lung were considerably higher than those in other tissues.     

一条在人睾丸中高表达的新的环指蛋白(RNF20)

从人胎脑构建的cDNA文库中,通过大规模测序筛选的方法获得一条新基因,该基因蛋白序列含有1个环指（RING finger)基序及2个进核信号（nuclear localization signal NLS)序列,钭此基因命名为RNF20（RING Finger 20),用放射杂交细胞系（Radial hybrid RH)方法定位此基因在人染色体的9q22上,Northern杂交表明,其mRNA在睾丸组织中高表达,用小鼠睾丸做原位杂交表明,小鼠的同源基因在精原细胞及初级精母细胞中高表达,未成年小鼠睾丸中的精原细胞没有发达,提示RNF20可能参与精子发生的调控作用。