Distraction osteogenesis (DO) represents a highly effective method for addressing significant bone defects; however, it necessitates a long treatment period. Exosomes are key mediators of intercellular communication. To investigate their role in the angiogenesis and osteogenesis of DO, we established a canine mandibular DO model with a bone defect (BD) group as the control. Higher levels of angiogenesis were observed in the regenerating tissue from the DO group compared to those from the BD group, accompanied by earlier osteogenesis. Proteomic analysis was performed on circulating exosomes at different phases of the DO using a data-independent acquisition method. Data are available via ProteomeXchange with the identifier PXD050531. The results indicated specific alterations in circulating exosome proteins at different phases of DO, reflecting the regenerative activities in the corresponding tissues. Notably, fibronectin 1 (FN1), thrombospondin 1 (THBS1), and transferrin receptor (TFRC) emerged as potential candidate proteins related to the angiogenic response in DO. Further cellular experiments validated the potential of DO-associated circulating exosomes to promote angiogenesis in endothelial cells. Collectively, these data reveal previously unknown mechanisms that may underlie the efficacy of DO and suggest that exosome-derived proteins may be useful as therapeutic targets for strategies designed to improve DO-related angiogenesis and bone regeneration.

The relationship between interleukin (IL)-10 and IL-6 gene polymorphisms and squamous cell carcinoma (SCC) has been demonstrated but with inconsistent conclusions. The aim of this study was to evaluate the potential associations of IL gene polymorphisms and the SCC risk. PubMed, Cochrane Library, Web of Science, China National Knowledge Infrastructure, China Biomedical Database, WanFang, and China Science and Technology Journal Database databases were searched for articles reporting the correlations of IL-10 and IL-6 gene polymorphisms with the SCC risk. Odds ratio and 95% confidence interval were calculated using Stata Version 11.2. Meta-regression, sensitivity, and publication bias were analyzed. False-positive reporting probability and Bayesian measure of the false-discovery probability were used to explore the credibility of the calculation. Twenty-three articles were included. The IL-10 rs1800872 polymorphism showed a significant correlation with the SCC risk in the overall analysis. Studies pooled by ethnicity revealed that the IL-10 rs1800872 polymorphism reduced the SCC risk in the Caucasian population. The results of this study suggest that the IL-10 rs1800872 polymorphism may confer a genetic susceptibility to SCC, particularly oral SCC, in Caucasians. However, the IL-10 rs1800896 or IL-6 rs1800795 polymorphism was not significantly associated with the SCC risk.

目的:研究旨在通过检测microRNA(miRNA)的表达特征来预测口腔鳞状细胞癌(OSCC)患者的存活率.方法:从TCGA数据库下载397名OSCC患者的表达谱数据及相应的临床信息.通过生物信息学的方法分析OSCC与正常组织间的差异表达的miRNA,使用Cox回归分析和其他生物信息学方法筛选预后相关的miRNA.并应用采用Kaplan-Meier分析和、受试者工作特征(ROC)曲线分析评估所筛选的miRNA作为预后评估指标的可信度.结果:通过生物信息学的方法分析获得差异表达的miRNA363个,其中上调的miRNA197个,下调的miRNA166个(PDR＜0.05).通过单变量COX回归分析发现84个miRNA的表达与患者预后显著相关,将其中P＜0.001的11个miRNA进一步进行多变量COX回归分析,其中4个miRNA(has-miR-30e、has-miR-337、has-miR-6507、has-miR-1251)纳入了风险评估模型.根据多因素COX分析的回归系数,构建由4个miRNA组成的预后风险评估模型,根据风险评分将OSCC患者分为高风险组和低风险组.Kaplan-Meier生存曲线表明高风险组生存率显著低于低风险组生存率(p=1.026e-05),构建的ROC曲线下面积AUC为0.669,C-index为0.63.结论:4个miRNA,has-miR-30e、has-miR-337、has-miR-6507及has-miR-1251的组合可以作为预测OSCC患者预后的潜在标志物.

BACKGROUND:Long noncoding RNAs (lncRNAs) are essential and critical components of signal and transduction, regulating the intracellular microenvironment. Serum exosomes (SEs) are involved in rearranging the intercellular functional lncRNAs, which may also play a role in oral squamous cell carcinoma (OSCC). The function of lncRNAs at the transcription level in SEs of patients with OSCC is partially understood.MATERIALS AND METHODS:The lncRNA expression profiles were examined derived from SEs from patients with OSCC with lymph node metastasis (OSCC-LNM), OSCC with no LNM (OSCC-NLNM), postoperative metastasis and recurrence OSCC (rOSCC) and healthy controls (HCs). Bioinformatics analysis was used to analyse differentially expressed lncRNAs (DE lncRNAs) and a total of 150 subjects were enrolled for RT-PCR verifications. The correlations of four lncRNAs and clinicopathologic factors, biochemical indexes were evaluated. MAGI2-AS3 and CCDC144NL-AS1 were overexpressed or silenced in oral cancer (OC) cells. The proliferation, invasion, and migration were evaluated to investigate the effect of MAGI2-AS3 and CCDC144NL-AS1 on the development of OSCC. The related proteins of PI3K-AKT-mTOR signal pathway were also detected.RESULTS:The expressions of the lncRNAs, namely MAGI2-AS3 and CCDC144NL-AS1, were significantly upregulated in rOSCC and OSCC-LNM. MAGI2-AS3 was overexpressed in cancer tissue compared to other control groups. AC109587.1 and AC010978.1 were significantly associated with the clinical stage, and CCDC144NL-AS1 was significantly associated with aging. MAGI2-AS3 and CCDC144NL-AS1 might promote cell proliferation, invasion, and migration in OSCC cells by regulating the PI3K-AKT-mTOR pathway.CONCLUSIONS:our results suggest that MAGI2-AS3 an d CCDC144NL-AS1 may have clinical applications in the treatment of OSCC.

Objectives: This study was designed to identify a messenger RNA (mRNA) expression signature to predict survival in patients with oral squamous cell carcinoma (OSCC). Methods: mRNA expression profiles were integrated with clinical data from 280 samples, including 19 normal tissues and 261 OSCC tissues in The Cancer Genome Atlas. We identified differentially expressed mRNAs (DEmRNAs) between the OSCC and normal tissue samples and developed a novel mRNA-focused expression signature using a Cox regression analysis and other bioinformatic methods. The prognostic value of this signature was evaluated by Kaplan-Meier analysis, multivariable COX regression, and receiver operating characteristic (ROC) curve analysis. Protein-protein interaction (PPI) network, gene ontology, and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were performed to predict the function of the DEmRNAs. Signature-related mRNAs were analyzed by gene set enrichment analyses (GSEA) and validated by quantitative real-time polymerase chain reaction (qRT-PCR) in 20 paired OSCC and adjacent healthy tissues. Results: We identified a novel 5-mRNA expression signature (HOXA1, CELSR3, HIST1H3J, ZFP42, and ASCL4) that could predict patient outcomes in OSCC. The risk score based on the signature was able to separate OSCC patients into high- and low-risk groups that showed significantly different overall survival (p < 0.001, log-rank test). The signature was further validated as an effective independent prognostic predictor of OSCC by multivariate Cox regression analysis (hazard ratio = 3.747, confidence interval: 2.279-5.677, p < 0.001) and ROC curve of the third year (area under the curve = 0.733). Functional analysis demonstrated that the key hub genes in the PPI network were mainly enriched in cell division, cell proliferation, and the p53 signaling pathway. GSEA results showed that the 5 mRNAs were significantly enriched in mismatch repair, DNA replication, and the NOTCH signaling pathway. Finally, qRT-PCR results showed that the 5 mRNAs were upregulated in OSCC tissue in agreement with the predictions from our bioinformatics analysis. Conclusions: We identified a novel 5-mRNA signature that could predict the survival of patients with OSCC and may be a promising biomarker for personalized cancer treatments.

生物技术是全球发展最快的高新技术之一,其技术过程复杂,基础理论知识体系庞大且复杂难懂.对多数学生来说,生物技术知识较为晦涩,不容易掌握,长期以往容易对这门课程丧失兴趣.而"投其所好"教学法,即以学生本来的兴趣为基础,将专业知识与其兴趣爱好相融合,辅以现代多媒体教学手段,能够培养其对生物技术专业的学习兴趣,同时使其更加容易接受和掌握相关的专业知识,对于学生专业知识的学习与学业发展具有重要的作用.

Genome stability is critical for the normal development of preimplantation embryos, as DNA damages may result in mutation and even embryo lethality. Anti-silencing factor 1A (ASF1A) is a histone chaperone and enriched in the MII oocytes as a maternal factor, which may be associated with the maintenance of genome stability. Thus, this study was undertaken to explore the role of ASF1A in maintaining the genome stability of early mouse embryos. The ASF1A expressed in the preimplantation embryos and displayed a dynamic pattern throughout the early embryonic development. Inhibition of ASF1A expression decreased embryonic development and increased DNA damages. Overexpression of ASF1A improved the developmental potential and decreased DNA damages. When 293T cells that had been integrated with RGS-NHEJ were co-transfected with plasmids of pcDNA3.1-ASF1A, gRNA-NHEJ, and hCas9, less cells expressed eGFP, indicating that non-homologous end joining was reduced by ASF1A. When 293T cells were co-transfected with plasmids of HR-donor, gRNA-HR, hCas9, and pcDNA3.1-ASF1A, more cells expressed eGFP, indicating that homologous recombination (HR) was enhanced by ASF1A. These results indicate that ASF1A may be associated with the genome stability maintenance of early mouse embryos and this action may be mediated by promoting DNA damage repair through HR pathway.

Rotavirus (RV) is the leading cause of viral gastroenteritis in neonates and VP6 protein has been discussed as a potential candidate vaccine. CRISPR/Cas9 was the latest generation of gene editing tools that can mediate the site-specific knock-in of exogenous genes, providing strong support for the expression of recombinant proteins. Here, seeking to design a rotavirus vaccine that would be suitable for both mammary-gland-based production and milk-based administration, rabbit β-casein ( CSN2 ) locus was chosen as the target site to integrate the VP6 gene. The efficiency of inducing mutations in different target sites of rabbit CSN2 locus was analyzed and g4 site seems to be the best one to generate mutations (g4 72.76 ± 0.32% vs g1 30.14 ± 1.93%, g2 38.53 ± 0.75%, g3 52.26 ± 1.16%, P < 0.05 ). We further compared the knock-in efficiency through cytoplasmic injection of two group mixtures (containing 100 ng/μL Cas9 mRNA or Cas9 protein, 20 ng/μL sgRNA4, and 100 ng/μL donor vector) in rabbit zygotes, though the Cas9 mRNA group induced an HDR efficiency as high as 20.0% ± 2.6% than Cas9 protein group (10.3% ± 3.1%), 37.5% of the knock-in events were partial integration in the target site, when Cas9 protein used in the CRISPR/Cas9 system, all of the positive blastocysts showed completely integrated, results showed that the use of Cas9 protein is better than Cas9 mRNA to integrate the correct exogenous gene into the target site. Moreover, the transgenic rabbit that harbored correct integration of VP6 gene was obtained using Cas9 protein group and was used to produce an experimental milk-based rotavirus vaccine. Our research provides a novel strategy to produce rotavirus subunit vaccine and make a foundation for building broader milk-based vaccine protection against other pathogens.

Coat color is of great importance in animal breed characteristics; it is not only a significant productive trait but also an indispensable economic trait, especially in the rabbit industry. In the present study, the relationship between melanocortin 1 receptor (MC1R) genotypes and coat color phenotypes was observed in five rabbit breeds with popular coat colors that are present in China. These breeds comprised the Lianshan black rabbit (BR), Fujian yellow rabbit (YR), New Zealand white rabbit (WR), Gray Giant rabbit (GR), and Checkered Giant rabbit (CR), which were firstly determined, and the results showed that GR had an E allele; WR, CR, and BR had a 6-bp in-frame deletion (c.281_286del6, ED allele); and YR had a 30-bp deletion (c.304_333del30 E allele). To explore the feasibility of obtaining a novel rabbit coat color through the mutation of MC1R with the CRISPR/Cas9 system, two single-guide RNAs (sgRNAs) were designed for the MC1R gene, and the editing efficiency was confirmed by injection of rabbits’ zygotes. Unlike the donor rabbits whose coat color was originally black, two novel pale-yellow-coated rabbits were generated in the founders. A total of six novel MC1R gene deletions were identified in the two founder rabbits, in which the longest deletion was more than 700 bp. The histological hematoxylin-and-eosin (H&E) staining results indicated that eumelanin amounts were absent in hair follicles of MC1R-knockout (KO) rabbits, when compared with that of donor BR. In addition, the messenger RNA (mRNA) levels of some key downstream genes in the MC1R pathway were all downregulated in MC1R-KO rabbits compared with BR and YR. These results further indicate that loss-of-function MC1R contributed to blocking the synthesis of eumelanin and created a novel pale-yellow coat color in the MC1R-KO rabbits, and gene editing technology may be a useful tool to generate novel phenotypes in rabbit breeding.

Myostatin (MSTN), a protein encoded by growth differentiation factor 8 (GDF8), is primarily expressed in skeletal muscle and negatively regulates the development and regeneration of muscle. Accordingly, myostatin-deficient animals exhibit a double-muscling phenotype. The CRISPR/Cas9 system has proven to be an efficient genome-editing tool and has been applied to gene modification in cells from many model organisms such as Drosophila melanogaster, zebrafish, mouse, rat, sheep, and human. Here, we edited the GDF8 gene in fibroblasts and embryos of Debao pig and swamp buffalo using the CRISPR/Cas9 system. The CRISPR/Cas9-mediated mutation efficiency in fibroblasts was as high as 87.5% in pig and 78.9% in buffalo. We then obtained single-cell clones with mutations at the specific sites of the GDF8 gene by screening with G418 in fibroblasts of pig and buffalo. In addition, the frequencies of Cas9/gRNA-mediated mutations were at 36 and 25% in the intracytoplasmic sperm injection embryos of pig and in vitro fertilization embryos of buffalo, respectively. Our work demonstrates that the Cas9/gRNA system is a highly efficient and fast tool for genome editing in cultured cells and embryos of Debao pig and swamp buffalo. These results can be helpful for the establishment of a new animal strain that can generate more meat.

试验旨在探讨单精子胞浆内注射(intracytoplasmic sperm injection,ICSI)法生产猪体外受精卵和应用猪ICSI受精卵进行胞质注射生产转基因胚胎的可行性.首先对比了猪体外受精(in vitro fertilization,IVF)受精卵与ICSI受精卵的胚胎发育效率;然后观察了猪ICSI受精卵的双原核形成时间及效率,对精子注射到胞质后6～18 h分6个时间段进行地衣红染色,对比精子进入卵胞质后的状态及原核形成;最后对猪IVF受精卵受精后8～10 h及ICSI受精卵受精后12～14 h进行EGFP-N1质粒(20 ng/μL)胞质注射,观察胚胎发育效率及转基因效率.结果表明,ICSI受精卵的胚胎发育率(卵裂率89.4％和67.9％、囊胚率36.5％和16.1％)显著优于IVF组(P＜0.05),适合用于猪的体外受精卵试验;猪ICSI受精卵双原核在精子注射到卵胞质后12～14 h形成,双原核形成率为54.90％,显著高于其余5个试验组(P＜0.05);ICSI受精卵胞质注射组胚胎卵裂率(86.2％和66.3％)、囊胚率(30.0％和13.6％)及转基因效率(18.5％和0)均显著高于IVF受精卵胞质注射试验组(P＜0.05).本试验结果为采用ICSI受精卵进行胞质注射生产转基因猪的研究奠定了基础.

Polled trait of goat is important economic trait in livestock production,goat polled inter sex syndrome (PIS) influenced polled phenotype and intersex phenotype of goats.PIS sequence locats in a approximately 11.7 kb nucleotide sequence in chromosome 1.This experiment was aimed to study the effect of PIS sequence on polled phenotype in Guanzhong dairy goats (3 horned goats,3 polled goats).The PIS whole sequence in Guanzhong dairy goats were amplified using four pair of primers (PIS1-1,PIS1-2,PIS2 and PIS3),the whole absence of PIS sequence was detected using the fifth primer (PIS whole).The sequencing result was assembled,analyzed and annotated.The result showed that the PIS sequence full length was 12.814 kb,and the BLAST alignment result was correct.PIS whole primer didn't amplify any srtipes,which showed that there was no a total deletion of 11.7 kb in Guanzhong dairy goats.SeqMan software analysis result showed that there was a total of 30 polymorphic sites,wihch were likely to connect with polled/horned phenotype in Guanzhong dairy goats.The annotation result of concatenation sequence showed that there were one tRNA encoding locus,two microsatellites,three microRNA encoding loci,L1-EN conserved domain,RT-nLTR-like conserved domain,also find a CpG island,one pair of inverted repeats,and two tandem repeats.This experiment result had a significant reference value for studying the molecular mechanism of polled phenotype in Guanzhong dairy goats.

The aim of this study was to clone the JHDM2A gene of porcine and study the expression of JHDM2A gene in porcine ovary.In this study, we cloned the porcine JHDM2A gene and constructed its eukaryotic expression vector, the expression of JHDM2A gene in porcine ovarian tissue was also analyzed.The results showed that the cloned CDS length of porcine JHDM2A gene was 3 945 bp, which encoded 1 315 amino acids.The results of multiple amino acid sequence comparison showed that the porcine JHDM2A shared 93.5%, 94.7%, 94.7% and 93.8% homologous compared with those of Bubalus bubalis, Bos taurus, Ovis aries and Homo sapiens, respectively.Phylogenetic tree analysis indicated that the JHDM2A gene was highly conservative in the evolutionary process.The pLVX-IRES-ZsGreen1-JHDM2A eukaryotic expression vector was constructed, and clear green fluorescent signal was observed when the plasmid was transfected into HEK293T cells by liposome method.The immunohistochemical results showed that the JHDM2A protein was expressed in porcine follicle of different development stages.The results showed that the porcine JHDM2A gene sequences was cloned, and the JHDM2A protein was highly expressed in porcine ovary, indicating that its function might be closely related to the development of porcine follicular.

In this study ,through cloning buffalo MBD3 gene and analyzing the biological informa‐tion of MBD3 gene sequence ,and constructing the expression vector of buffalo MBD3 ,to provide a basis for the function research of buffalo MBD3 gene on embryo development and iPCSs .The total RNA was extracted from buffalo fresh ovary ,and MBD3 gene was amplified and sequenced , the sequence was systemically analysed with bioinformatics techniques .And the MBD3 CDS was cloned into the pEGFP‐C1 vector .Then the recombinant plasmid pEGFP‐C1‐MBD3 was trans‐ferred into the HEK293T cells and buffalo fetal fibroblasts (BFF) ,the expression was analyzed by RT‐PCR ,Western blotting and fluorescence microscope .The results showed that 898 bp of MBD3 gene fragment including whole 774 bp CDS was cloned and sequenced ,and encoded 257 amino acids .The multiple sequence alignment and analysis of phylogeny tree showed that MBD3 gene was highly conserved in the process of evolution .Especially the MBD domain ,the MBD do‐main of buffalo MBD3 gene shared 100% of similar nucleotide sequence with Bos taurus ,and shared 97% of similar nucleotide sequence with Homo sapiens ,Sus scrof a and Pan troglodytes . The recombinant plasmids pEGFP‐C1‐MBD3 were transferred into HEK293T cells and BFF ,fluo‐rescence observation ,RT‐PCR and Western blotting were used to analyze the expression of buffa‐lo MBD3 .The results suggested that the expression vector of buffalo MBD3 gene was successfully constructed .T he study laid the foundation for the function research of MBD 3 on embryo develop‐ment and inducing of iPCSs .

Cytoplasmic microinjection (CI) of foreign gene into in vivo fertilized zygotes has emerged as a useful tool for transgenic pig production. In the current study, we investigated factors affecting transgenic efficiency and developmental potential of parthenogenetic (PA) and in vitro-fertilized (IVF) porcine embryos produced by CI. These factors included adding of RNase inhibitor, DNA or RNA concentration, injection time, and different structures of plasmids. Our results showed that adding of 1–4 U/μL of RNase inhibitor did not have negative effect on development potential of CI-PA embryos, and RNase inhibitor injection significantly increased EGFP expressing rate of CI-PA embryos. High injection DNA concentration and long injection interval after PA significantly reduced blastocyst formation. Different molecular structures such as DNA or RNA affected CI-PA embryos development, and RNA had little harmful effect on pig’s early embryonic development. EGFP expression rate of CI-IVF embryos was improved following the increase of foreign DNA concentration, but blastocyst formation rate was decreased. Injection time after IVF did not show any significant difference on embryonic development, but longer interval resulted in a significantly lower EGFP expressing rate. Cas9 mRNA and myostatin (GDF-8) sgRNA co-injection indicated that the mutation rate of CI-IVF group was significantly higher than that of CI-PA. The CI-IVF-generated embryos were then transferred to six recipient pigs, but no live piglets were obtained. The following pronuclear formation assessment showed more than 76.1% IVF zygotes were polyspermy. These results demonstrate that CI-PA and CI-IVF were effective methods for production of transgenic pig embryos. However, polyspermic fertilization and poor quality of porcine IVF blastocysts are still the main problem of resulting in pregnancy failure.

试验旨在对沼泽型水牛ALK3基因进行克隆、生物信息学分析,并对其在水牛组织中的表达规律进行系统研究.根据GenBank中已公布的牛ALK3基因序列设计特异性引物,应用RT PCR方法扩增、克隆获得目的基因片段;应用生物信息学方法分析和预测了水牛ALK3的遗传进化及蛋白质的理化性质、二级和三级结构;并应用QRT-PCR技术对ALK3基因在水牛组织中的表达进行了差异分析.结果表明,水牛ALK3基因编码区全长1 599 bp,共编码532个氨基酸.多重序列比较分析显示,水牛ALK3核苷酸序列与牛、绵羊、猪、马、人和小鼠相应序列的同源性分别为98％、96％、95％、93％、94％和91％.系统进化树分析显示,ALK3基因在不同物种以及进化的过程中具有高度保守性.对ALK3蛋白质的分析表明该蛋白呈弱碱性,有信号肽,细胞亚定位于胞膜上,存在丝氨酸/苏氨酸激酶和GS等结构.定量分析结果显示,ALK3在水牛生殖脊、心脏、肝脏、颗粒细胞、肺脏、卵丘细胞、肾脏、下丘脑、垂体、大脑等15种组织或细胞中有不同程度的表达,其中卵巢中表达量最高,垂体、肺脏和睾丸次之,卵丘细胞表达量最低.本研究成功克隆了沼泽型水牛ALK3基因,并研究了其在不同水牛组织细胞中的表达规律,为阐明其在水牛繁殖过程中的功能及转基因载体构建中的应用研究奠定了理论基础.

为了克隆水牛ATG5、LC3基因序列,并分析自噬相关基因在水牛各组织中的表达情况,试验采用RT-PCR技术克隆了水牛自噬相关基因ATG5、LC3序列,并对其CDS序列进行了生物学信息分析,同时利用QRT-PCR方法,对ATG3、ATG5、ATG6、ATG7、ATG12、LC3、BECN1 7个基因在胎水牛心脏、肝脏和肌肉等9种组织、成年水牛卵巢中的mRNA表达水平进行了分析.结果表明:克隆得到水牛ATG5基因序列980 bp,其中包括CDS序列828 bp,预计编码275个氨基酸;水牛LC3基因mRNA序列全长499 bp,包括CDS序列378 bp,预计编码125个氨基酸;所分析的7个自噬相关基因在10种水牛组织中均有不同程度的表达.

为了阐明水牛ALK5基因在水牛卵泡发生过程中的作用,了解其在水牛不同组织中的表达模式,试验采用了RT-PCR及QRT-PCR方法对ALK5基因进行了研究,参考已公布的GenBank中牛ALK5基因序列,设计特异性引物应用RT-PCR方法扩增克隆获得目的基因片段;应用生物信息学方法,系统分析和预测了水牛ALK5基因的遗传进化、蛋白质的理化性质、二级和三级结构,并应用QRT-PCR技术对ALK5基因在水牛组织的表达进行了差异分析.结果表明:水牛ALK5基因的编码区全长为1 512 bp,共编码503个氨基酸;水牛ALK5核苷酸序列与牛、绵羊、猪、人和小鼠相应序列的相似性分别为99％、97％、95％、93％和90％,结合系统进化树分析结果推测ALK5基因在不同物种以及进化的过程中具有高度保守性;该蛋白呈弱碱性,有信号肽,细胞亚定位于胞膜上,存在lowcomplexity和GS结构,呈典型跨膜蛋白结构;水牛ALK5基因在肺脏、垂体、下丘脑组织中不表达,在水牛大脑、肝脏、骨骼、卵巢、肌肉和心脏等12个组织细胞中具有不同程度的表达,其中卵巢中表达量最高,心脏和肾脏次之,肌肉表达量最低.说明试验成功地克隆了沼泽型水牛ALK5基因,其具备典型的TGF-β受体家族结构,表达具有一定的组织特异性.