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مفهوم التعدبل الوراثي
أضرار وفوائد التعديل الوراثي
تقنيات التعديل الوراثي للنباتات والحيوانات
ماهو التعديل الوراثي?
إن التعديل الجيني والتعديل الوراثي هما مصطلحان متردافان لمفهوم واحد ، حيث ان العوامل الوراثية تكون محملة على الجينات داخل الخلايا الحية ، والمقصود بالتعديل الوراثي هو نقل الجينات ذات المواصفات المرغوبة من كائن لاخر بهدف تحسين الجودة او زيادة الانتاج او مكافحة الامراض والافات.
جميع الكائنات الحية تحتوي على جينات في حمضها النووي الريبوزي المنزوع الأكسجين (DNA ) . وهذه الجينات عبارة عن تعليمات كيميائية لبناء وحفظ الحياة العلماء لهم القدرة على تغيير خصائص الكائن عن طريق تعديل الجينات. يتم تعزيز وزيادة حجم العضلة وزيادة قدرة النبات وحيوانات المزرعة على مقاومة الأمراض وذلك عن طريق تعديل الجينات . الهندسة الجينية تتمتع بهذه الاحتمالات . ولكن تكنولوجيا الجينات ما زالت في بداية التطور . نسبة قليلة جدا من المحاصيل الغذائية عدلت جينيا باستخدام التكنولوجيا الحديثة . لا يوجد في العالم حتى الآن محاصيل معدلة جينيا يتم زراعتها تجاريا باستخدام جينات منقولة من الحيوانات أو البشر.
كيف يتم تعديل كائن وراثيا ؟
التعديل الوراثي هو نقل مورثة من كائن إلى آخر … و هذه العملية تتم في مراحل …
1_النسخ العكسي
2_ تلميم المورثة المعزولة
3_دمج المورثة في الخلية العائلة
4_زرع الخلايا العائلة في مخمرات صناعية
التعديل الوراثي عند النباتات
التعديل الوراثي هو إدخال صفات وراثية جديدة على صنف ما من النباتات باستخدام التقنيات البيولوجية ( الحيوية ) يحسن من نوعية و جودة المنتج الزراعي . فمثلا تضاف جينات بعض النباتات سريعة النمو إلى النباتات بطيئة النمو بهدف زيادة كمية الإنتاج .
هذا و قد كان أول نبات تجرى عليه عملية التعديل الوراثي هي الطماطم و يتمثل هدف التعديل في إطالة فترة نضجها و عدم فسادها في وقت سريع .
الفرق بين النباتات المعدلة وراثيا و بين النباتات العادية .
الهدف من زراعة النباتات العادية و النباتات المعدلة وراثيا واحد وهو إنتاج أنواع محسنة وذات إنتاج أكبر لكن الاختلاف هو في الطرق التي يتم زراعة هذه النباتات بها فالنباتات العادية هي النباتات التي تزرع بصورة عادية و تكون طبيعية 100% حتى و إن هجنت مع أصناف أخرى بهدف تحسين الإنتاج . أما النباتات المعدلة وراثيا فيتم باستخدام أحدث التكنولوجيا المتوصل إليها في عالم الهندسة الوراثية و نقل الجينات المطلوب نقل صفاتها من صنف إلى آخر بواسطة بعض أنواع البكتيريا أو بما يعرف بقاذف الجينات البيولوجي .
أمثلة على التحسينات التي يقوم بها التعديل الوراثي .
1) إنتاج محصول أكبر من النباتات التي لم تعدل وراثيا
2)إنتاج أنواع من الأرز مثلا تحتوي على البروتينات الموجودة في الفول .
فوائدة و مضار
فوائده
كما ذكرت أن فوائده تعتمد على إنتاج سلالات ذات فوائد غذائية أكبرأو تحويل الخصائص الجينية لبعض النباتات مثل إنتاج البطيخ بدون بذر كما يزيد من مقاومة النبات للحشرات
فمثلا نبتة القطن تتم مهاجمتها من قبل حشرة تدعى دودة القطن التي قد تهلك محصول كامل من القطن فتم تطوير نوعية محسنة من نبتة القطن تحتوي على مضادات لهذه الحشرة الأمر الذي يضمن عدم مهاجمة هذه الحشرة للقطن و بالتالي سلامة محصول القطن .
أضراره
يقول الخبراء أن أضرار هذه النباتات المعدلة وراثيا لا تظهر على الفرد مباشرة بعد أكلها بل تتأخر أعراضها إلى فترات طويلة نسبيا قد تمتد إلى سنوات . و لا تزال الكثير من أضرار هذه النباتات محل جدل بين المنتجين و الوكالات الصحية .
و لم تثبت دراسة صحية أكيدة حتى الآن أن هذه النباتات لها ضرر واضح على الإنسان . الأمر الذي جعل المنتجين يزيدون من إنتاجها دون إجراء اختبارات الأمان الكافية لمعرفة هل هي مناسبة للإستحدام الآدمي أو لا . مما جعل الصراع على أشده بين الوكالات الصحية و منتجي هذه الأصناف . و قد تم التوصل أخيرا إلى اتفاق بين هذه الوكالات و المنتجين سيتم تطبيقه بعد فترة قصيرة يقضي بوجوب كتابة عبارة يحتوي على مواد معدلة وراثيا أو عبارة : لا يحتوي على المواد المعدلة وراثيا .
و بهذا يكون الخيار للمشتري في شراء منتجات تحتوي على مواد معدلة وراثيا غير مأمونة من الناحية الصحية أو بقائه على المنتجات الزراعية العادية .
نظرة مهندسو الجينات وعلماء الهندسة الوراثية
يقول مهندسو الجينات بأنه لا يوجد أي دليل على وجود غذاء معدل جينياً في المحلات تسبب في تأثيرات مرضية ، ويعتمدون على نقطة توجد لدى جميع الأغذية المصنعة بعيداً عن التعديل الجيني وهي أن جميع المكونات الأساسية تمت معالجتها للتخلص من المواد السامة أو الغير مرغوب فيها، وهذا ما يوجه الغذاء المعدل جينياً ليكون آمناً أكثر من الغذاء المصنع.
-إن التعديل الجيني يمكن أن يسبب ظهور أنواع جديدة من الحساسية ، وكذلك التهجين التقليدي للنباتات يمكن أن يتسبب في المثل، هكذا يقول مهندسو الجينات ولكن التقنية الجديدة يمكن أن تمنح إمكانية تعديل المشكلة بعيداً عن الغذاء.
-إن استخدام الــ DNA من الفيروسات والبكتيريا التي في النباتات يشكل القليل من الخطورة تحديداً لأننا لسنا نباتات . القرنبيط أو الزهرة ( Cauliflowers ) تتغذى طبيعياً بفيروس يستخدم بكثرة في المختبرات لأغراض التعديل الجيني والكثير يأكلون الملايين من الخضار بدون شكوى من أي أعراض صحية . بالإضافة إلى ذلك فإن التعديل الجيني يمكننا من تطوير الطعم ، القوام والشكل ، فترة الصلاحية وكذلك القيمة الغذائية ، وبإمكاننا زيادة مقدار الفيتامينات في الخضراوات والفواكه / إدخال مضادات السرطنة والتقليل من الدهنيات المضرة للصحة .
يدافع المهندسون الحيويين أيضاً بأن تكنولوجيا التعديل الجيني سوف تتطلب كيماويات قليلة لها سمية قليلة ولها القدرة على التحلل السريع والبقاء في التربة عوضاً عن انتقالها إلى مجاري الأنهار . وفوق ذلك لها القدرة على إنتاج محصول وفير , وهذا بدوره يقلل الضغط على المساحات المزروعة الغير محدودة . ويعمل العلماء على دراسة إمكانية تعديل النباتات لتنتج بلاستيك ووقود غير ضار للبيئة عوضاً عن المنتجات التي تعتمد على مشتقات النفط.
لا شك أن هذا “الاختراع” الجيني يعد بارقة أمل ونقطة تحول كبيرة في سبيل مكافحة ظواهر الفقر وسوء التغذية والجوع التي تودي بحياة الملايين سنويا ، فقد ظهرمؤخرا العديد من الاكتشافات التي تحمل في طياتها آمالا مشرقة تبرز بعض الآثار الإيجابية للنباتات والحيوانات المعدلة جينيا على النظم البيئية والحياة الإنسانية، كنجاح فريق علمي أميركي في تحوير نبات الخردل الهندي جينيا من أجل تنقية التربة الزراعية من أحد أخطر العناصر المعدنية السامة وهو عنصر السيلنيوم الذي يتسبب في إصابة الإنسان بالسرطان وأمراض أخرى.
ولم تقف إنجازات الهندسة الوراثية عند حد المساهمة في التخلص من السموم وملوثات التربة، بل أمتد الأمر لأوجه أخرى عديدة منها المساهمة في القضاء على واحدة من أكبر المشكلات البيئية التي تواجه العالم ألا وهي مشكلة التصحر حيث يمكن بواسطة تلك النباتات إعادة زراعة أو تشجير ملايين الأفدنة من الأراضي القاحلة التي أتى عليها الجفاف أو التصحر.
وليست هذه الأمثلة في الواقع إلابعض من إنجازات وفضائل عديدة لا يمكن إنكارها على الهندسة الوراثية ومنتجاتها، لكن هذا على أي حال لا ينفض كامل الغبار عن باقي المنتجات المعدلة وراثياً، فقد يقولون غدا أنه يمكن أن نحصل على الفيتامينات والعناصر الضرورية التي نريدها في فاكهة أو ثمرة واحدة هي التفاحة العجيبة …!
النظرة المتخوفة للنقاد ومنظمات حماية البيئة والإنسان
هناك مخاوف وشكوك متزايدة تتعلق بجدوى استقدام كثير من النباتات والحيوانات المعدلة جينياً تتجلى في عدة نقاط ومن أكثر من وجهة نظر :
1- ضيق المعرفة العلمية في هذا المجال : يحتج المعترضون والناقدون بأننا لا نعلم عن الطريقة التي تعمل وتتفاعل بها الجينات بما فيه الكفاية لنضمن نتيجة أي تعديل جيني ، ويخافون من أن يؤدي التعديل الجيني إلى وجود مواد سامة أو مسببة للحساسية مثلا، والمعارضون للتعديل الجيني ينتقدون بشدة استخدام الفيروسات والبكتيريا النباتية في التعديلات الجينية ويخشون من احتمال أن تسبب أمراضاً جديدة قد لايمكن السيطرة عليها ، كما وعارضوا استخدام الجينات المقاومة للمضادات الحيوية التي يستخدمها العلماء للتأكد من نجاح التعديلات التي أدخلوها في النبات.
ويحتج المعارضون أيضاً على أن الجينات المقاومة للمضادات الحيوية يمكن أن تنتقل إلى الحيوانات الدقيقة التي تسبب الأمراض وبالتالي نصبح غير قادرين على استخدام المضادات الحيوية للقضاء على هذه الكائنات الدقيقة.
2-عدم اتباع المتطلبات والاحتياطات اللازمة عند التطوير أو التعامل مع الكائن المعدل جينياً، وأهمها ضمان التحكم في انتشار ذلك الكائن واتخاذ جميع التدابير لمنع تسربه إلى البرية أو الحيلولة دون انتقال جيناته المعدلة لأنواع أخرى وهو ما يعرف باسم التلوث الجيني.
وليس في هذا أدنى مزايدة، بل هو احتمال قائم إذ أن هناك من الشواهد والأمثلة ما يشير إلى حدوثه فعلاً، منها ما حدث في الولايات المتحدة الأميركية مؤخراً عندما تم استزراع أحد أنواع البطاطس المعدلة وراثيا القادرة على مقاومة نوعية معينة ضارة من الخنافس، لكن ما حدث هو أن الضرر أمتد فعلا لنوعية ثانية من الخنافس تبين لاحقاً أنها ذات فائدة حيوية ونفع لعدة محاصيل زراعية أخرى.
هذا ما ترتب على ظهور واحد فقط من المنتجات المعدلة جينياً، ترى هل تم التأكد فعلاً من سلامة عشرات المنتجات المعدلة الأخرى على البيئة المحيطة وعلى عناصرها؟. سؤال عويص أشك أن لدى علماء الهندسة الوراثية إجابة حاسمة عليه بعد. ولا يعني هذا سوى تزايد احتمالات تسرب عدد كبير من المنتجات المعدلة جينيا للأسواق أو البرية دون القيام بدراسات كافية أو دون تمام التأكد من كونها آمنة أو صحية بالنسبة للبيئة والإنسان.
3-الخوف المتزايد من الاستغلال السيء للكائنات المعدلة جينيا وذلك من قبل فئة ضالة من العلماء أو المستغلين، لأنه إذا كانت غالبية العلماء من الفئة الجادة والمخلصة التي تحرص على اتباع الأصول العلمية وتلتزم بالأطر الأخلاقية والمهنية، فهناك أيضاً، وإن كانت قلة، فئة أخرى لديها رغبة ملحة في الشهرة الذائعة وربما الكسب السريع حتى ولو كان ذلك على حساب كل المعايير الأخلاقية المعروفة.
والحقيقة أن هناك من المؤشرات والأمثلة السيئة ما يكفي لإثراء تلك المخاوف، وهي تدل أولا وقبل أي شيء على مغالاة الإنسان ومزايدته في استخدام أو إنتاج نباتات أو حيوانات مهندسة جينيا لا داعي لها ولا جدوى من استقدامها.
فما جدوى إنتاج نوع مماثل من نجيلة الحدائق تشع ضوءا في الظلام أو سلالة جديدة من الأرانب أو القطط الفسفورية المتوهجة. ما معني هذا وما مغزاه؟! لا شك أنه عبث بيوتكنولوجي جامح وفوضى جينية منفلتة لا فائدة منها سوى تغذية سفه الرفاهية المفرطة للإنسان ، وهذا عن العبث الجيني المعلن، لكن ماذا عن المخفي؟، ماذا عن الملفات والأبحاث الخفية في مئات المعامل والمختبرات السرية؟! ألا يمكن أن يقود هذا إلى ظهور كائنات ممسوخة ذات صفات تركيبية شاذة وضارة؟ ثم ألا يمكن أن يقود هذا إلى كارثة بيئية تضر بالأرض والحياة الإنسانية القائمة عليها؟.
4-اختلال النظام البيئي : تخشى الجماعات المهتمة بالبيئة من أن الجينات المهندسة في المحاصيل قد تتمكن من الهروب وأن تنقل إلى فصائل نباتية أخرى تؤثر بها تأثيراً سلبياً،وخصوصا الجينات التي لها القدرة على مقاومة المبيدات الحشرية والنباتية، فهم يعتقدون أن تسرب بعض الجينات يمكن أن ينتج ظهور بذور قوية (Superweeds ) واختفاء بعض فصائل الحشرات والطيور وتحطيم السلسلة الغذائية . وهم يتهمون شركات التكنولوجيا الحيوية بأنها تحاول أن تقيد علم الزراعة من خلال محاولة إجبار المزارعين التعامل مع خيارات قليلة وشراء البذور المعدلة جينياً والمواد الكيماوية المصاحبة لها .
يمكن إنتاج نبات أو حيوان منيع إزاء مرض معين في إطار برنامج ”تقليدي” للإكثار، أي من خلال التهجين مع سلالات الأقارب المنيعة، ثم الاصطفاء، فالتهجين الرجعي. كما يمكن القيام بذلك عبر إدخال مورِّثة تمنح المناعة المطلوبة بالاعتماد على الهندسة الوراثية. وفي حين أن النوعين الناتجين عن كلا هذين الأسلوبين سيتمتعان بالمناعة، فإن النوع الثاني فحسب يعتبر كائناً حياً معدلاً وراثيا. والجديد في هذا هو قدرة العلماء على كشف التُمامة الوراثية ودراسة مورِّثات الكائن الحي ثم استخدام هذه المعلومات في تغيير الكائن الحي، بل وحتى نقل المورِّثات من كائن حي الى آخر بعيد عنه كل البعد في سلم التطور. وهذا بالضبط هو مبعث الجدل.
ويقول بيان منظمة الأغذية والزراعة حول التكنولوجيا الحيوية ”إن المنظمة تقر بأن الهندسة الوراثية تتمتع بالقدرة على زيادة الإنتاج والقدرة الإنتاجية في قطاعات الزراعة، والغابات، ومصايد الأسماك”. ويمضي البيان قائلاً ”إنها يمكن أن تزيد الغلات في المناطق الحدية في البلدان العاجزة الآن عن إنتاج ما يكفي من غذاء لإطعام شعوبها”. غير أن البيان يضيف بأن المنظمة ”تدرك أيضاً القلق المنبعث بسبب المخاطر المحتملة التي تخلقها بعض جوانب التكنولوجيا البيولوجية. وهذه المخاطر تندرج في فئتين رئيسيتين هما: الآثار على صحة الإنسان والحيوان، والعواقب البيئية”.
وتتيح الأدوات الجديدة فرصاً جديدة أيضاً لمعالجة المشكلات الزراعية التي عجزت التقنيات التقليدية عن حلها. وفي العادة فإن تطوير الكائنات الحية المعدَّلة وراثياً يتم على يد المصالح التجارية. وباستثناء بضعة حالات فإن هذه التقانة لم تعد بالفائدة على صغار المزارعين حتى الآن.
الهندسة الوراثية الحيوانية ::
و تهدف إلى تحسين عناصر الإنتاج الحيواني، و زيادة إنتاج المستحضرات المناعية والتشخيصية والعلاجية للحيوان والبرمجة الوراثية لتحسين الأسماك. لقد تطورت التكنولوجيا الحيوية تطوراً كبيراً ابتداءً من بدء تعامل الإنسان مع الكائنات الحية على أسس علمية راسخة بالطرق التقليدية إلى أن وصلت الآن إلى تطورها الحديث، حيث بلغ تعامل الإنسان مع المادة الحية أقصى درجات الدقة فيما يعرف بتطعيم الجينات ونقلها من كائن إلى آخر.
تعتبر الإنجازات العلمية الهائلة التى سطعت فى منتصف القرن الحالي من اكتشاف طبيعة المادة الوراثية، كذلك اكتشاف آليات ابتناء البروتين … اللبنات الأولى فى تطور التكنولوجيا الحيوية لتصل إلى مفهومها الحالى … حيث نتج عن هذه الاكتشافات تطور مذهل فى علوم الوراثة، مما أدى إلى تغير كبير فى الكثير من طرق تناول حقائق العلوم الأساسية (النبات والحيوان وغيرهما)، وكذلك تطور الأساليب البحثية المستخدمة فى التكنولوجيا الحيوية بمجالاتها المختلفة، وأخيراً ظهور التكنولوجيا الحيوية المتقدمة.
وقد تطورت التكنولوجيا الحيوية تطوراً جذرياً منذ أوائل السبعينيات واتسعت وتشعبت نشاطاتها بدرجة كان لها آثار متعاظمة فى اقتصاديات العالم. ثم جاءت ثورة الإنزيمات، ومن بينها إنزيمات البلمرة وإنزيمات القطع المتخصصة وإنزيمات النسخ العكسية وغيرها. والتى أسرعت من تداول وتناول المادة الوراثية فى مخطط متكامل للوصول إلى هدف معين. ويدور هذا التطور حول حجر زاوية فريد من نوعه، وهى أن الوراثيين بإمكانهم الآن وضع المادة الوراثية على مائدة العمليات لتصبح مطوعة للتغيير كمًّا ونوعاً، بحيث تحذف منها مقاطع أو يضاف إليها ويعاد صياغتها بحيث تعبر عن ذاتها بطريقة جديدة، وهو ما يسمى بالتطعيم الجينى .. وهو اتجاه معاصر فى علوم البيولوجيا تبلور خلال العقود القليلة الماضية، تنحصر مهمته فى برمجة أشكال من المناهج المبتكرة، وبمعنى آخر التحكم فى الصفات بكم وكيف يفوق كثيراً كل ما أعطته كافة الطرق الأخرى كالانتخاب والتهجين والتطفير.
لقد تعرضت مجالات الإنتاج الحيوانى لثورتين .. أولهما هى الثورة الخضراء الأولى: التىأسفرت عن استخدام الوسائل التقليدية فى تحسين عناصر الإنتاج الحيوانى، ثم الثورة الخضراء الثانية: التى اعتمدت على التعامل الدقيق مع المادة الحية فيما يعرف بالهندسة الوراثية لإضافة الجينات الخاصة بزيادة الإنتاج وإنتاج المستحضرات المناعية والتشخيصية والعلاجية للحيوان والبرمجة الوراثية لتحسين الأسماك.
النسخ والاستنساخ(Cloning):
يشتهر بين الناس كلمة الاستنساخ نظرا لارتباطها بخلق الكائنات أو إنشاء نُسخ منها.و لكن بالمصطلح الطبي فان كلمة نسخ أو استنساخ تعنى عملية إنشاء صورة طبقا الأصل من المادة التي يراد نسخها.و قد يكون النسخ لقطعة من الدي إن أي أو نسخ كائن حي متكامل.و لا شك أن لغتنا العربية تفرق بين كلمة نسخ و استنساخ و لكننا سوف نستخدم كلمة نسخ أو استنساخ في حديثنا لنعني نفس الشيء. و في كلمة استنساخ باللغة العربية تعني ( Cloning)وينتج عنه نسخة أو مستنسخ (Clone).
عندما قام الدكتور… و فريقه العلمي بنشر (Nature 385, 810-13, 1997 )خبر استنساخ النعجة “دولي” في احد مختبرات اسكتلندا ( مختبر روزيلين ) عام 1997 زاد اهتمام العالم بموضوع الاستنساخ و زاد الفضول العلمي في الحديث عن استنساخ الإنسان و فجر ذلك الخبر الكثير من التحفظات الدولية من كثير من المراكز الدينية و العلمية على الجانب الأخلاقي من عملية استنساخ الإنسان.
و بعد ذلك اخبر أصبحت كلمة استنساخ تستخدم بين العامة في الحديث عن عملية خلق نسخة أخرى من الحيوان أو الإنسان و بذلك بدأ البس بين الكثيرين في معنى هذه الكلمة.و لا شك فان العلماء كانوا و مازالوا يستعملون هذه الكلمة في الإشارة إلى عملية صنع نسخة من أي مادة وراثية و ليس بالضرورة خلق أو نسخ كائن حي بالكامل. و لذلك فالعلماء يقسمون الاستنساخ أو النسخ إلى 3 أنواع :
1- نسخ أو استنساخ القطع من الدي إن أي عن طريق الهندسة الوراثية و بما يعرف بتهجين الدي إن أي ( Recombinant DNA technology ) .
2- الاستنساخ التكاثري أو الجنسي Reproductive cloning
3- الاستنساخ العلاجيTherapeutic cloning نسخ أو استنساخ القطع من الدي إن أي عن طريق الهندسة الوراثية.
إن ما يهتم به العلماء في باب الاستنساخ هو نسخ قطع من الدي إن أي كانت هذه القطع عبارة عن جين(مورث)أو جميع الجينوم(أي كل الدي إن أي الموجود في الكائن الحي).و اشهر العمليات التي تجرى هي نسخ قطعة من الدي إن أي. و يحتاج العلماء للقيام بنسخ القطع لأنهم يحتاجون إلى كمية كبيرة من هذه النسخ و ذلك لندرة استخلاصها في كل مرة من داخل الخلية و ذلك لوجود التعقيدات الإنشائية للكروموسومات .و على سبيل المثال فان الجين المنتج لسلسة بيتا في الهيموجلوبين و المعروف بمورث ببيتا جلوبين( Beta-Globin Gene) يمثل فقط 0.00005% من حجم الدي إن أي الكلي في الخلية(و الذي يتراوح ب 3بلايين قاعدة نووية).كما أن الجين العملاق و المعروف بجين الدستروفين( Dystrophin Gene ) و الذي يتراوح حجمه بال2.5 ميقابيز (2.5Megabases ) لا يمثل أكثر من 0.08% من الحجم الكلي للدي إن أي في الخلية.و لذلك فان العلماء يحتاجون إلى إجراء نسخ لهذه الجينات أو القطعة من الدي إن أي لكي يتسنى لهم التعامل بها و إجراء التجارب عليه
1- تصميم القطع مهجنة من الدي إن
يعتمد النسخ باستخدام الخلايا الحية على قدرة القطعة المراد نسخها على الانقسام أو التكاثر الذاتي عندما توضع داخل الخلية الحية.و لا شك أن قطع الدي إن أي العادية ليس لديها القدرة على التكاثر الذاتي و لذلك فان العلماء قاموا بتجاوز هذا الأمر بان ادخلوا القطعة التي يريدون نسخها في ناقل من النواقل( Vectors ) المعروفة بقدرتها على التكاثر الذاتي(راجع موضوع النواقل).و بغض النظر عن نوع الناقل فان طريقة إدخال قطعة الدي ان أي المراد نسخها إلى الناقل تقريبا واحدة.و هذه الخطوات ببساطة كما يلي:
1- بعد أن يتم تحديد القطعة المراد نسخها يضاف إليها إنزيم قاطع محدد و ليكن مثلا إنزيم ( أ )فيقوم هذا الإنزيم بقطع الدي إن أي في مكان محدد حسب التسلسل النووي.
2- يضاف نفس الإنزيم للناقل و الذي يقوم بقطعة أيضا في نفس التسلسل النووي.
3- تضاف القطع المراد نسخها بعد قطعها بالإنزيم القاطع إلى الناقل المقطّع. فتتداخل التسلسلات النووية بين الناقل و بين قطع الدي إن أي المراد نسخها. فنشاء من ذلك قطعة مهجنة من الناقل و بداخله القطعة المراد نسخها.و ترتبط قطعة الدي إن أي البلازميد من أطرافها برابطة هيدروجينية و هي رابطة ضعيفة لذلك يضاف إنزيم يسمى ليقيز أو اللاصق(Ligase ) لكي يحول الترابط بين قطعة الدي إن أي و التناقل إلى رابطة قوية(Covalent Bond )
إن أكثر لناقلات استخداما هي البلازميد و لكن يمكن استخدام الفيج أو الياك أو أي ناقل أخر.و الذي يحدد نوع الناقل المراد استخدامه هو في العادة كبر القطعة المراد استنساخها.ففي حالة القطع الصغيرة يستخدم البلازميد أو الفيج بينما يستخدم الياك أو الباك في حالة القطع الكبيرة.
2- نقل القطعة المهجنة و التي هي بداخل الناقل إلى خلية حية
في الغالب تستعمل البكتيريا خاصة النوع المعروف الايكولي(E.Coli) في عملية الزراعة و ذلك لسهولة إدخال الناقل إليها، و إلى سرعة انقسامها (تنقسم البكتريا تقريبا كل 20 دقيقة)، إضافة إلى توفر طرق الاختيار خاصة التي تعتمد على خاصة الحماية من المضادات الحيوية.و يدخل البلازميد أو الفيج تلقائيا إلى داخل البكتيريا بينما الناقلات الأخرى تحتاج إلى مساعدة ،و في العادة بتغيير تركيز الأملاح المحيطة بالبكتيريا أو تعرض إلى نبضة كهربائية لكي يسمح الجدار المحيط بالبكتيريا بدخول الناقلات.و من طبيعة البكتريا إنها تنقسم تلقائيا و بشكل سريع و كذلك البلازميدات.
3- اختيار المستعمرات البكتيرية التي تحتوي على الناقل و القطعة المهجنة
مع تكاثر الخلايا البكتيرية و تكاثر البلازميد التي بداخلها ينتج لدينا أعداد كثيرة من المستعمرات البكتيرية و بها البلازميد المهجن.و لكن قد يكون في داخل الطبق الذي زرع فيه البكتريا بعض البكتريا التي لا تحتوي على البلازميد المهجن و لكي يمكن التعرف على البكتيريا التي تحتوي على البلازميد المهجن فنه في العادة يقام باستعمال ناقلات عليها جينات واقية من المضادات الحيوية، كالجين الواقي من المضاد الحيوي امبيسيلين أو لاتترسيلين و غيرها. و بذلك فالمضاد الحيوي سوف يمنع تكاثر أي خلية بكتيرية لا تحتوي على البلازميد المهجن و الذي علي الجين الواقي من المضاد الحيوي.
4- استخلاص القطع المهجنة و استخراج الدي ان أي منها بكميات كبيرة. بعد أن يُتعرف على المستعمرات التي تحتوي على البلازميد المهجن فانه يمكن نقلها إلى طبق جديد و يحافظ عليها و تغذى لكي تستمر بالتكاثر.و هذه البكتيريا يكون فيها أعداد كثيرة من البلازميد و بذلك تنتهي عملية النسخ.و يستفاد من هذه القطع المنسوخة في القيام بالمزيد من البحوث أو التجارب عليها كان يقام مثلا إنتاج مكتبة من الدي إن أي أو محاولة استنتاج التسلسل النووي للقطعة.كما يمكن تحوير هذه العملية بحيث يحتوي البلازميد على قطعة من سي دي إن أي(cDNA) بدل و من ثم تحوير المراحل الأخيرة من الزراعة لإنتاج بروتين بدلا من الدي إن أي. و هذه الطريقة هي التي تستعمل في إنتاج بعض الهرمونات كهرمون النمو .
الاستنساخ التكاثري (Reproductive cloning ) استنساخ الكائنات الحية بالكامل
يعرف الاستنساخ التكاثري أو الجنسي بأنه إنتاج لكائن حي له نفس المادة الوراثية( Nuclear DNA ) لكائن حي أخر المنسوخ منه. لقد قام الفريق العلمي بمختبر روزلين بعملية استنساخ جنسي في عملية استنساخ للنعجة دولي.و تعرف هذه العملية أيضا ًبنقل نواة الخلية الجسمية (somatic cell nuclear transfer” (SCNT ). و بشكل مبسط نقل نواة من خلية من خلايا الجسم غير الجنسية أي غير التي توجد في المبيض (في الأنثى )و من خلايا الخصية(في الذكر).و الخلية التي استعملت لاستنساخ دولي كان من خلايا الثدي لنعجة أخرى.و من ثم أخذت أيضا بويضة من المبيض و قام العلماء من التخلص من النواة التي بداخل تلك البويضة ثم قاموا بزرع النواة التي أخذوها من ثدي في داخل البويضة. ثم قاموا بصعق تلك البويضة بالكهرباء لكي ينشطوا عملية الانقسام. و بعد أن بدأت هذه البويضة في الانقسام قاموا بغرزها داخل رحم نعجة و بعدها نما الجنين في الرحم فأصبح”بإذن الله” نعجة كاملة.
علميا فان دولي (أو أي حيوان أو إنسان )يستنسخ بهذه الطريقة ليس في الحقيقة نسخة مطابقة للام أو الأب الذي اخذ منه النواة. فهناك بعض من المادة الوراثية موجود خارج النواة و هو بالتحديد موجود في داخل البويضة التي أزيل منها النواة.و هذه المادة الوراثية موجودة على جسيمات صغيرة تسمى بالميتوكوندريا (Mitochondria ). و مع أن الميتوكوندريا مصنع هام للطاقة إلا انه يكثر فيها الطفرات مع تقدم العمر وقد يكون لها علاقة بالهرم.
الاستنساخ العلاجي Therapeutic cloning
و يقصد بذلك استنساخ كائنات حية لأخذ خلايا جذعية(Stem Cells) و لا يسمح لها للوصول إلى تخليق كائن حي كامل. و أهمية هذه الخلايا تنبع في قدرة هذه الخلايا في إنتاج أي خلايا أو أعضاء كالكلية و الكبد و الخلايا الدموية و التي يرجى في استخدامها علاج الكثير من الأمراض التي لا يوجد لها علاج شافي. و لقد قامة إحدى الشركات العلمية في ولاية ماسيشيوستز بالولايات المتحدة الأمريكية (Advanced Cell Technologies ) في شهر نوفمبر من عام 2001 بالإعلان عن محاولة ناجحة لاستخلاص خلايا جذعية من أجنة مستنسخة و ذلك بعد أن قامة باستخدام 8 بويضات بشرية تم تفريغها من نواها ثم زرع بداخلها نوى خلايا من الجلد.و لقد نجحوا في إنتاج خلايا جذعية من بويضة واحدةبينما فشلت البويضات السبع.
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