綿羊TYRP基因快速進化以適應(yīng)海拔變化
發(fā)布時間：2018-11-23 21:01
【摘要】：為研究高海拔綿羊群體強紫外線輻射適應(yīng)性的分子機制,為綿羊群體間毛色多樣性形成機制研究提供理論參考。本研究以分布于不同海拔高度的綿羊群體:高原型藏羊、歐拉型藏羊、山谷型藏羊、烏珠穆沁羊、蘇尼特羊、灘羊、廣靈大尾羊、小尾寒羊和湖羊作為試驗材料,選擇與黑色素合成相關(guān)的光保護基因TYRP1和TYRP2,采用混合樣測序技術(shù)檢測綿羊群體內(nèi)這兩個基因的SNP位點,采用直接測序的方法對其進行分型,并對這些位點的基因頻率及基因的單倍型頻率與海拔高度進行關(guān)聯(lián)分析;同時對12個物種的TYRP1和TYRP2基因系統(tǒng)發(fā)生關(guān)系、基因相似性和選擇方向進行系統(tǒng)分析,并且用Lositan軟件對9個綿羊群體內(nèi)檢測到的TYRP基因的SNP位點進行受選擇狀況分析;利用生物信息學(xué)的方法對TYRP蛋白的結(jié)構(gòu)進行預(yù)測分析,分析TYRP基因變異位點和TYRP蛋白結(jié)構(gòu)的關(guān)系,進一步闡述TYRP的基因結(jié)構(gòu)與其蛋白質(zhì)功能之間的關(guān)系,為中國綿羊的選育工作和遺傳資源保護利用提供理論參考。研究結(jié)果如下:1.混合樣測序在綿羊群體內(nèi)TYRP基因中共檢測到35個SNP位點,其中TYRP1基因檢測到21個位點,TYRP2基因中檢測到14個位點。對這些位點的等位基因頻率與各綿羊群體所在海拔高度進行關(guān)聯(lián)分析發(fā)現(xiàn),TYRP1基因中有5個SNP位點的等位基因頻率和海拔高度顯著相關(guān),分別是g.11121CT、g.11124TC、g.26549CG、g.26682CT和g.26730CA,其中g(shù).26682CT位點所在密碼子編碼的氨基酸為綿羊TYRP1蛋白磷酸化位點,g.26730CA位點所在密碼子編碼的氨基酸位于綿羊TYRP1蛋白的跨膜區(qū)域,g.11121CT位點的等位基因頻率在高海拔群體分別與中低海拔兩個綿羊群體間差異顯著,同時在TYRP1基因中還有3個單倍型頻率與海拔高度顯著相關(guān);TYRP2基因中沒有檢測到與海拔變化顯著相關(guān)的變異。由此推測,TYRP1基因的變異在綿羊毛色形成中可能發(fā)揮著重要的作用。2.對綿羊TYRP基因編碼的蛋白的生物信息學(xué)預(yù)測分析得到,綿羊TYRP1基因編碼537個氨基酸,1-24位氨基酸區(qū)間為信號肽區(qū)域,跨膜區(qū)域有兩段,分別是7-24和479-501,磷酸化位點有27個,二級結(jié)構(gòu)預(yù)測結(jié)果顯示其螺旋結(jié)構(gòu)(Helix)占14.71%,延鏈(Strand)占5.59%,環(huán)狀結(jié)構(gòu)(Loop)占79.70%,86-465位氨基酸為TYRP1蛋白的功能域;綿羊TYRP2基因編碼517個氨基酸,1-23位氨基酸區(qū)間為信號肽區(qū)域,跨膜區(qū)域為469-491,磷酸化位點有23個,二級結(jié)構(gòu)預(yù)測結(jié)果顯示其螺旋結(jié)構(gòu)占17.02%,延鏈結(jié)構(gòu)占3.48%,環(huán)狀結(jié)構(gòu)占79.50%,82-456位氨基酸為TYRP2蛋白的功能域。TYRP1和TYRP2蛋白的結(jié)構(gòu)具有很高的相似性。3.TYRP基因在12個物種間正選擇檢驗發(fā)現(xiàn),TYRP1基因受到顯著的正選擇作用,TYRP2基因主要受到純化選擇的作用。9個綿羊群體間35個SNP位點正選擇檢驗結(jié)果顯示,TYRP1基因中有5個SNP位點受到正選擇作用,一個位點受到平衡選擇。正選擇位點分別是g.10941CG、g.11055CG、g.11168CA、g.26717CT和g.26790AC,其中g(shù).10941CG位點所在密碼子編碼的氨基酸位于TYRP1蛋白的信號肽區(qū)域,g.26717CT位點所在密碼子編碼的氨基酸在綿羊TYRP1蛋白的跨膜區(qū)域,g.11055CG位點所在的密碼子編碼的氨基酸位于綿羊TYRP1蛋白氨基酸序列的第46位,該氨基酸位點在物種間基因選擇方向檢驗結(jié)果中同樣受到顯著的正選擇作用,兩者檢測的結(jié)果相一致;TYRP2基因中檢測到3個位點受到平衡選擇的作用,分別是g.28506CA、g.28629CT和g.40404AC,其中g(shù).28629CT位點所在密碼子編碼的氨基酸位于綿羊TYRP2蛋白的功能域,g.40404AC位點所在密碼子編碼的氨基酸為綿羊TYRP2蛋白的跨膜區(qū)域。由此可以推測,TYRP1基因受到的正選擇作用和TYRP2基因受到的平衡選擇的作用在綿羊群體適應(yīng)不同海拔高度的不同紫外線輻射強度的環(huán)境中可能發(fā)揮著重要作用。
[Abstract]:In order to study the molecular mechanism of the adaptability of the high-altitude sheep population to the strong ultraviolet radiation, a theoretical reference is provided for the study of the formation mechanism of the wool-color diversity among the sheep. The research is to distribute the sheep group with different altitude: the plateau-type Tibetan sheep, the Euler-type Tibetan sheep, the valley-type Tibetan sheep, the black-pearl-muqin sheep, the Suits sheep, the beach sheep, the wide-and-long-tailed sheep, the small-tail cold sheep and the lake sheep as the test materials, selecting the light protection genes TYRP1 and TYRP2 related to the synthesis of melanin, detecting the SNP sites of the two genes in the sheep population by adopting a mixed sample sequencing technology, and typing the SNP sites of the two genes in the sheep population by adopting a direct sequencing method, and carrying out the correlation analysis on the gene frequency and the single-fold frequency of the gene and the altitude of the genes, and simultaneously carrying out system analysis on the relationship between the TYRP1 and TYRP2 gene systems of the 12 species, the gene similarity and the selection direction, and the structure of the TYP protein is predicted and analyzed by using the bioinformatics method, the relationship between the TYP gene mutation site and the TYP protein structure is analyzed, The relationship between the gene structure and its protein function of TYP is further elaborated, which provides a theoretical reference for the breeding of Chinese sheep and the protection of genetic resources. The results of the study are as follows: 1. A total of 35 SNP sites were detected in the TYP gene in the sheep population by the hybrid-like sequencing, of which the TYRP1 gene was detected at 21 sites and 14 sites were detected in the TYRP2 gene. The allele frequency of these sites was correlated with the altitude of each sheep population, and the allele frequency and altitude of the five SNP loci in the TYRP1 gene were significantly related, g. 11121CT, g. 11124TC, g. 26549CG, g. 26682CT and g. 26730CA, respectively. wherein the amino acid encoded by the codon of the g. 26682CT site is the phosphorylation site of the sheep TYRP1 protein, the amino acid encoded by the codon at the position where the g. 26730CA site is located is located in the transmembrane region of the sheep TYRP1 protein, There were also 3 single-fold frequencies in the TYRP1 gene that were significantly related to the altitude; there was no variation in the TYRP2 gene that was significantly related to altitude change. It is suggested that the variation of the TYRP1 gene may play an important role in the formation of the color of the wool. The bioinformatics prediction of the protein encoded by the sheep TYP gene is obtained, 537 amino acids are encoded by the sheep TYRP1 gene, the 1-24 position amino acid interval is a signal peptide region, two sections of the transmembrane region are respectively 7-24 and 479-501, and the phosphorylation sites are 27, The results of the two-stage structure show that the helix structure (Helix) is 14.71%, the strand (Strand) is 5.59%, the loop structure (Loop) is 77.9%, the 86-465 amino acid is the functional domain of the TYRP1 protein, the sheep TYRP2 gene encodes 517 amino acids, the 1-23 position amino acid interval is the signal peptide area, the transmembrane region is 469-491, The results of the two-stage structure prediction showed that the helical structure was 17. 02%, the structure of the extension chain was 3.48%, the ring-like structure was 70.50%, and the 82-456-position amino acid was the functional domain of the TYRP2 protein. TYRP1 and TYRP2 protein have a very high similarity. Five SNP sites in the TYRP1 gene are under positive selection and one site is balanced. The positive selection sites are g. 10941CG, g. 11055CG, g. 11168CA, g. 26717CT and g. 26790AC, wherein the amino acid encoded by the codon of the g. 10941CG site is located in the signal peptide region of the TYRP1 protein, the amino acid encoded by the codon at which the g. 26717CT site is located is in the transmembrane region of the sheep TYRP1 protein, g. the amino acid encoded by the codon at which the 11055CG site is located is located at the 46th position of the amino acid sequence of the sheep TYRP1 protein, and the amino acid site is also subjected to a significant positive selection effect in the test result of the inter-species gene selection direction, and the result of the two detection is consistent; The three sites detected in the TYRP2 gene were selected by the balance selection, g. 28506CA, g. 28629CT and g. 40404AC, where g. 28629CT site's codon-encoded amino acid is located in the functional domain of the sheep TYRP2 protein, and the amino acid encoded by the codon of the g. 40404AC site is the transmembrane region of the sheep TYRP2 protein. It is thus possible to speculate that the positive selection action of the TYRP1 gene and the effect of the balanced selection of the TYRP2 gene play an important role in the environment where the sheep population is adapted to different ultraviolet radiation intensity at different altitudes.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S826
本文編號：2352685
[Abstract]:In order to study the molecular mechanism of the adaptability of the high-altitude sheep population to the strong ultraviolet radiation, a theoretical reference is provided for the study of the formation mechanism of the wool-color diversity among the sheep. The research is to distribute the sheep group with different altitude: the plateau-type Tibetan sheep, the Euler-type Tibetan sheep, the valley-type Tibetan sheep, the black-pearl-muqin sheep, the Suits sheep, the beach sheep, the wide-and-long-tailed sheep, the small-tail cold sheep and the lake sheep as the test materials, selecting the light protection genes TYRP1 and TYRP2 related to the synthesis of melanin, detecting the SNP sites of the two genes in the sheep population by adopting a mixed sample sequencing technology, and typing the SNP sites of the two genes in the sheep population by adopting a direct sequencing method, and carrying out the correlation analysis on the gene frequency and the single-fold frequency of the gene and the altitude of the genes, and simultaneously carrying out system analysis on the relationship between the TYRP1 and TYRP2 gene systems of the 12 species, the gene similarity and the selection direction, and the structure of the TYP protein is predicted and analyzed by using the bioinformatics method, the relationship between the TYP gene mutation site and the TYP protein structure is analyzed, The relationship between the gene structure and its protein function of TYP is further elaborated, which provides a theoretical reference for the breeding of Chinese sheep and the protection of genetic resources. The results of the study are as follows: 1. A total of 35 SNP sites were detected in the TYP gene in the sheep population by the hybrid-like sequencing, of which the TYRP1 gene was detected at 21 sites and 14 sites were detected in the TYRP2 gene. The allele frequency of these sites was correlated with the altitude of each sheep population, and the allele frequency and altitude of the five SNP loci in the TYRP1 gene were significantly related, g. 11121CT, g. 11124TC, g. 26549CG, g. 26682CT and g. 26730CA, respectively. wherein the amino acid encoded by the codon of the g. 26682CT site is the phosphorylation site of the sheep TYRP1 protein, the amino acid encoded by the codon at the position where the g. 26730CA site is located is located in the transmembrane region of the sheep TYRP1 protein, There were also 3 single-fold frequencies in the TYRP1 gene that were significantly related to the altitude; there was no variation in the TYRP2 gene that was significantly related to altitude change. It is suggested that the variation of the TYRP1 gene may play an important role in the formation of the color of the wool. The bioinformatics prediction of the protein encoded by the sheep TYP gene is obtained, 537 amino acids are encoded by the sheep TYRP1 gene, the 1-24 position amino acid interval is a signal peptide region, two sections of the transmembrane region are respectively 7-24 and 479-501, and the phosphorylation sites are 27, The results of the two-stage structure show that the helix structure (Helix) is 14.71%, the strand (Strand) is 5.59%, the loop structure (Loop) is 77.9%, the 86-465 amino acid is the functional domain of the TYRP1 protein, the sheep TYRP2 gene encodes 517 amino acids, the 1-23 position amino acid interval is the signal peptide area, the transmembrane region is 469-491, The results of the two-stage structure prediction showed that the helical structure was 17. 02%, the structure of the extension chain was 3.48%, the ring-like structure was 70.50%, and the 82-456-position amino acid was the functional domain of the TYRP2 protein. TYRP1 and TYRP2 protein have a very high similarity. Five SNP sites in the TYRP1 gene are under positive selection and one site is balanced. The positive selection sites are g. 10941CG, g. 11055CG, g. 11168CA, g. 26717CT and g. 26790AC, wherein the amino acid encoded by the codon of the g. 10941CG site is located in the signal peptide region of the TYRP1 protein, the amino acid encoded by the codon at which the g. 26717CT site is located is in the transmembrane region of the sheep TYRP1 protein, g. the amino acid encoded by the codon at which the 11055CG site is located is located at the 46th position of the amino acid sequence of the sheep TYRP1 protein, and the amino acid site is also subjected to a significant positive selection effect in the test result of the inter-species gene selection direction, and the result of the two detection is consistent; The three sites detected in the TYRP2 gene were selected by the balance selection, g. 28506CA, g. 28629CT and g. 40404AC, where g. 28629CT site's codon-encoded amino acid is located in the functional domain of the sheep TYRP2 protein, and the amino acid encoded by the codon of the g. 40404AC site is the transmembrane region of the sheep TYRP2 protein. It is thus possible to speculate that the positive selection action of the TYRP1 gene and the effect of the balanced selection of the TYRP2 gene play an important role in the environment where the sheep population is adapted to different ultraviolet radiation intensity at different altitudes.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S826
【參考文獻】
相關(guān)期刊論文 前1條
1 張琳西,王臻,郭樹忠,夏煒,李巍;中波紫外線對體外培養(yǎng)的人黑色素細(xì)胞的影響[J];中國美容醫(yī)學(xué);2001年03期
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