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تعیین هاپلوگروپ های ژنوم میتوکندری در اقوام کرد و یزد در ایران

تعداد 110صفحه در فایل word

کارشناسی ارشد رشته زیست شناسی- ژنتیک(MSC)

 

تعیین هاپلوگروپ های ژنوم میتوکندری در اقوام کرد و یزد در ایران

چکیده

زمینه و هدف: ژنوم میتوکندری در سلول های انسانی 16569 نوکلئوتید دارد. ناحیه کنترل موجود در ژنوم میتوکندری دارای نواحی HV1 و HV2 می باشد. سرعت ایجاد تنوع نوکلئوتیدی در این توالی ها ده برابر سریعتر از DNA کروموزومی می باشد و تنها از مادر به فرزندان به ارث می رسد. هدف از این تحقیق تعیین هاپلوگروپ ها وفراوانی آنها می باشد.

روش بررسی: در این مطالعه هاپلوتایپ های ناحیه HV1 و HV2 ژنوم میتوکندری 100 نفر از استان یزدو 100 نفر کرد مورد بررسی قرار گرفت. ابتدا 2 میلی لیتر خون محیطی در لوله های حاوی ضد انعقاد تهیه و پس از تخلیص DNA ژنومیک، دو ناحیه HV1 و HV2 تکثیر و تعیین توالی گردید. توالی ها توسط برنامه Chromas  با توالی مرجع مقایسه و پلی مورفیسم ها مشخص گشتند و بر اساس این تغییرات از طریق درخت فیلوژنتیک، هاپلوگروپ ها و فراوانی آن ها در این اقوام مشخص گردید. پس از آن در جهت تایید هاپلوگروپ ها، SNP های 7028 و 12308 ناحیه کنترل از طریق RFLP تعیین گردید.

یافته ها: در این اقوام هاپلوگروپ هایU,J,T ,L3a وH به صورت مشترک دیده شده است که هاپلوگروپ های U و T بالاترین فراوانی و هاپلوگروپ های M و IWXبافراوانی کمتری در شهر یزد دیده شده است. هاپلوگروپ J  با بالاترین فراوانی و هاپلوگروپ هایK و T با فراوانی کمتری در قوم کرد دیده شده است.

نتیجه گیری: این مطالعه در مقایسه با فراوانی هاپلوگروپ های سایر مناطق مخصوصا اروپا نشان داد که در این اقوام هاپلوگروپ های اوراسیای غربی غالب بوده است. اگر چه تعداد محدودی نیز از هاپلوگروپ های آسیایی و آفریقایی در این جمعیت ها دیده شده است.

واژگان کلیدی: ژنوم میتوکندری، هاپلوتایپ، هاپلوگروپ

فهرست مطالب

عنوان  

صفحه

فصل1. 1

کلیات   1

1-1. بخش اول.. 2

1-1-1. مقدمه. 2

1-1-2. دلیل انتخاب ژنوم میتوکندری… 2

1-1-3. مدل های پراکندگی جغرافیایی ژنوم میتوکندری… 3

1-2. بخش دوم: میتوکندری… 7

1-2-1. تاریخچه میتوکندری… 7

1-2-2. منشا و تکامل میتوکندری… 8

1-2-3. میتوکندری حاصل یک رویداد درون همزیستی… 8

1-2-4. ساختمان میتوکندری… 9

1-2-5. اندازه و شکل میتوکندری… 11

1-2-6. ژنوم میتوکندری… 12

1-2-7. وراثت مادری… 14

1-2-8. ژن های میتوکندریایی… 14

1-2-9. هماهنگی بین ژنوم میتوکندری و هسته. 16

1-2-10. همانندسازی ژنوم میتوکندری… 17

1-2-11. رونویسی و ترجمه ژنوم میتوکندری… 18

1-2-12. نقش زیستی میتوکندری… 18

1-2-13. هتروپلاسمی و هوموپلاسمی… 22

1-2-14. بیماری های ژنتیکی میتوکندریایی… 23

1-3. بخش سوم: هاپلوگروپ وهاپلوتایپ…. 24

1-3-1 تعریف هاپلوگروپ و هاپلوتایپ…. 24

1-3-2. طریقه نام گذاری و شکل گیری هاپلوگروپ…. 25

1-3-3. Mitochondria Eve. 27

1-3-4. Seven Daughters of Eve. 27

1-3-5. تعیین هاپلوگروپ ها 30

1-4. بخش چهارم: معرفی استان ها 35

1-4-1. استان کردستان.. 35

1-4-2. استان کرمانشاه 36

1-4-3. استان یَزد. 38

1-5. بخش پنجم: طرح پیشنهاده 41

1-5-1. طرح موضوع.. 41

1-5-2. بیان مسئله. 41

1-5-3. ضرورت انجام تحقیق.. 43

1-5-4. اهداف پژوهش….. 44

1-5-5. سوالات…. 44

1-5-6. فرضیه ها 44

1-5-7. تعاریف واژه 44

فصل2  45

مروري بر منابع مطالعاتي   45

فصل3   50

مواد و روش ها 50

3-1 نمونه گیری… 51

3-2 استخراج DNA به روش نمک اشباع.. 51

3-3. آماده سازی نمونه ها جهت انجام PCR.. 53

3-3-1 رسوب گذاری با اتانول.. 53

3-3-2. تعیین غلظت و کیفیت DNA توسط دستگاه Nanodrop. 54

3-4 واکنش زنجیره ای پلی مراز (PCR). 54

3-4-1. مواد مورد نیاز برای آزمایش PCR.. 54

3-4-2. توالی پرایمرها 55

3-4-3. شرایط بهینه شده برای PCR.. 55

3-4-4 الکتروفورز ژل آگارز به منظور بررسی کیفی محصول PCR.. 56

3-5. آماده سازی نمونه ها جهت توالی یابی… 57

3-6 روش RFLP.. 58

3-6-1 ترکیبات لازم جهت انجام RFLP.. 58

3-6-2. روش انجام RFLP.. 59

3-7. الکتروفورز ژل پلی اکریل آمید. 60

3-7-1. مواد و محلول های مورد نیاز جهت الکتروفورز ژل پلی اکریل آمید.. 60

3-7-2. روش تهیه ژل پلی اکریل آمید.. 61

3-7-3. بار گذاری نمونه ها روی ژل پلی اکریل آمید.. 61

3-8. رنگ آمیزی ژل پلی اکریل آمید.. 61

3-8-1. محلول های مورد نیاز جهت رنگ آمیزی ژل پلی اکریل آمید.. 61

فصل4   63

یافته ها 63

4-1. توالی یابی نمونه های مورد مطالعه. 64

4-2. هاپلوگروپ های تعیین شده در استان یزد. 65

4-3. هاپلوگروپ های تعیین شده در قوم کرد. 72

4-4. فراوانی هاپلوگروپ ها در جمعیت های مورد مطالعه. 77

فصل5  79

بحث   79

5-1. مقدمه. 85

5-2. مقایسه جمعیت های ایرانی… 87

5-3. مقایسه تغییرات نوکلوتیدی در جمعیت ها 88

5-4. مقایسه فراوانی هاپلو گروپ های جمعیت های مورد مطالعه و ایران.. 89

5-5. مقایسه نمونه های مورد مطالعه با کشور های مجاور. 90

5-6 مقایسه با کشورهای جهان.. 92

5-7. مقایسه هاپلوگروپ ها در جمعیت های مورد مطالعه. 93

5-8. درخت فیلوژنتیک قوم یزد. 95

5-9. درخت فیلوژنتیک قوم کرد. 96

5-10. فاصله ژنتیکی درون جمعیت یزد. 97

5-11. فاصله ژنتیکی درون جمعیت کرد. 97

فصل6  98

نتایج و پیشنهادات   98

6-1. نتیجه گیری… 99

6-2. پیشنهادات…. 99

فصل7   100

منابع  100

Abstract 106

 

فهرست اشکال

عنوان  

صفحه

شکل 1-1………………………………………………………………………………………………………………………………………………….. 4

شکل 1-2…………………………………………………………………………………………………………………………………………………. 5

شکل 1-3 ………………………………………………………………………………………………………………………………………………… 6

شکل 1-4…………………………………………………………………………………………………………………………………………………. 11

شکل 1-5………………………………………………………………………………………………………………………………………………….. 13

شکل 1-6…………………………………………………………………………………………………… ……………………………………………14

شکل 1-7…………………………………………………………………………………………………………………………………………………. 16

شکل 1-8…………………………………………………………………………………………………………………………………………………. 17

شکل 1-9………………………………………………………………………………………………………………………………………………… 19

شکل 1-10……………………………………………………………………………………………………………………………………………… 21

شکل 1-11……………………………………………………………………………………………………………………………………………… 34

شکل 1-12……………………………………………………………………………………………………………………………………………… 36

شکل 1-13……………………………………………………………………………………………………………………………………………… 38

شکل 1-14……………………………………………………………………………………………………………………………………………… 40

شکل 3-1………………………………………………………………………………………………………………………………………………………………60

شکل 4-1……………………………………………………………………………………………………………………………………………………………… 64

شکل4-2……………………………………………………………………………………………………………………………………………………………… 64

شکل 5-1………………………………………………………………………………………………………………………………………………… 93

شکل 5-2…………………………………………………………………………………………………………………………………94

شکل 5-3………………………………………………………………………………………………………………………………….94

شکل5-4………………………………………………………………………………………………………………………………….95

شکل 5-5…………………………………………………………………………………………………………………………………96

فهرست جداول

عنوان  

صفحه

جدول 1-1……………………………………………………………………………………………………………………………………………………….. ….26

جدول 1-2……………………………………………………………………………………………………………………………………….. 32

جدول 1-3……………………………………………………………………………………………………………………………………….. 33

جدول3-1 ………………………………………………………………………………………………………………………………………. 52

جدول 3-2 …………………………………………………………………………………………………………………………………….. 52

جدول3-3 ………………………………………………………………………………………………………………………………………. 53

جدول 3-4 …………………………………………………………………………………………………………………………………….. 55

جدول 3-5 ……………………………………………………………………………………………………………………………………. 55

جدول 3-6 …………………………………………………………………………………………………………………………………….. 56

جدول 3-7 ……………………………………………………………………………………………………………………………………. 58

جدول 3-8 ……………………………………………………………………………………………………………………………………. 59

جدول 4-1 …………………………………………………………………………………………………………………………………… 65

جدول4-2 …………………………………………………………………………………………………………………………………….. 72

جدول 4-3………………………………………………………………………………………………………………………………..77

جدول 4-4………………………………………………………………………………………………………………………………..78

جدول 5-1 ……………………………………………………………………………………………………………………………………. 86

جدول 5-2 ……………………………………………………………………………………………………………………………………. 87

جدول5-3 ……………………………………………………………………………………………………………………………………… 88

جدول5-4 ……………………………………………………………………………………………………………………………………… 89

جدول 5-5 ……………………………………………………………………………………………………………………………………. 91

جدول 5-6 ……………………………………………………………………………………………………………………… ……………..92

جدول 5-7 ……………………………………………………………………………………………………………………………….97

جدول5-8……………………………………………………………………………………………………………………………………97

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